MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry. Oligonol treatment reduced the overexpression of phospho-p38, phospho-ERK1/2, phospho-inhibitor of nuclear factor-kappa B (NF-κB), NF-κBp65, and NF-κBp65-induced inflammatory protein such as cyclooxygenase-2, inducible nitric oxide synthase, tumor necrosis factor-α, and interleukin-6. Specificity and sensitivity of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody. Aliquot and store at The activation of signal transduction pathways by growth factors, hormones and neurotransmitters is mediated through two closely related MAP kinases, p44 and … Keep on ice. Add protein A agarose (10–30 µl of 50% bead slurry). Microcentrifuge for 10 min at 4°C, 14,000 x g and transfer the supernatant to a new tube. Alternatively, cells may be stored overnight at 4°C in 1X PBS. Block specimen in Blocking Buffer for 60 minutes. NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water. To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). Supplied in SDS Sample buffer. Wash cells by centrifugation in excess 1X PBS to remove methanol. Vortex, then microcentrifuge for 30 sec. Molecular weight of MAP kinase (Erk2) protein is 42 kDa. View our alternatives for ab24157 or you can download the archived datasheet PDF from this page. While blocking, prepare primary antibody by diluting as indicated on product webpage in Antibody Dilution Buffer. Specificity: Phospho-ERK1/2 (Thr202/Tyr204) Antibody detects endogenous levels of ERK1/2 only when phosphorylated at Threonine 202/Tyrosine 204. 10X Cell Lysis Buffer: (#9803) To prepare 10 ml of 1X cell lysis buffer, add 1 ml cell lysis buffer to 9 ml dH2O, mix. As ERK1/2 has many substrates, different biological effects may occur when the signal proteins compete for the same target protein. The Ig molecule in monomeric form is a glycoprotein with a molecular weight … Lyse cells by adding 1X SDS sample buffer (100 µl per well of 6-well plate or 500 µl for a 10 cm diameter plate). This suggested that although ΔMEKK3:ER* can activate ERK1/2, JNK and p38, the ERK1/2 or ERK5 pathways were required for phosphorylation of Bim EL. Conduct a small-scale experiment if you are unsure. Molecular Weight: ~ 42,44 kDa: ... which was accompanied by sustained increases in phospho-ERK1/2 (p-ERK1/2) and phospho-Cdk5(S159) (p-Cdk5(S159)) in the spinal cord dorsal horn (SCDH). The kit includes enough antibody to perform two western blot experiments with each primary antibody. Heat a 20 µl sample to 95–100°C for 5 min; cool on ice. Store at -20°C Phospho-Erk1/2 Pathway Sampler Kit n 1 Kit (5 x 20 µl) Description: The Phospho-Erk1/2 Pathway Sampler Kit provides an economical means of evaluating multiple members of the Erk pathway as well as their activation state. individually using the catalog numbers listed below. NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly. - Find MSDS or SDS, a COA, data sheets and more information. However, note that some fluorophores (including PE and APC) are damaged by methanol and thus should not be added prior to permeabilization. Description: Rabbit polyclonal antibody to ERK1/2 (phospho-Y204) Specificity: p-ERK1/2 (Y204) pAb detects endogenous levels of ERK1 protein when phosphorylated at Tyr204, and ERK2 protein when phosphorylated at Tyr187. Trademark Information. NOTE: Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation if the epitope is disrupted by formaldehyde and/or methanol. Western blot analysis of extracts from Hela cells, untreated or treated with TPA (200nM), and NIH/3T3 cells, untreated or treated with PDGF (100ng/ml), using Phospho-Erk1/2 (Thr202/Tyr204) Antibody (upper) or Tubulin (lower). proteintech@ptglab.com, (+44) 161 839 3007 Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. 07-467 Anti-phospho -MAP Kinase1/2 (Erk1/2) IPFL07810 Immobilon-FL 7 x 8.4 cm PVDF 0.45 mm (sheet) 10/pk 06-642 Anti-phospho -MAP Kinase1/2 (Erk1/2) (Tyr180), polyclonal WBKLS0100 Immobilon Western Chemilu m HRP Substrate 100 mL 17-463 STAR ERK 1/2 ELISA Kit 17-373 Spray & Glow™ ECL WB Detection … 2 The Erk1/2 was inactivated by dephosphorylation through action of MAPK … This protocol is intended for immunoprecipitation of native proteins utilizing Protein A agarose beads for analysis by western immunoblot or kinase activity. Wash membrane-bound HRP (antibody conjugate) three times for 5 minutes in TBST. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 5 minutes and 200 nM PMA for 20 minutes.. PVDF membrane was probed … Supplied in SDS Sample buffer. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Alexa Fluor is a registered trademark of Life Technologies Corporation. Incubate with rotation overnight at 4°C. The Phospho(Thr202/Tyr204 Thr; 185/Tyr187) /Total ERK1/2 Assay is a sandwich immunoassay (Figure 1) M. SD provdei s a plaet tha thas been pre -coated with capture antibodies for phosphorylated ERK1/2 (Thr202/Tyr204; Thr185/Tyr187) and total ERK1/2on spatially distinct spots . Molecular weight of MAP kinase (Erk2) protein is 42 kDa. waste container. The kit includes enough antibody to perform two western blot experiments with each … No. Treat cells by adding fresh media containing regulator for desired time. Invitrogen Anti-Phospho-ERK1/ERK2 (Thr202, Tyr204) Polyclonal, Catalog # 36-8800. Cytometry Kit (Methanol) #13593, or - Find … Wash pellet five times with 500 µl of 1X cell lysis buffer. 027-87531627, Contact Us These findings suggest that ASP can prevent the janus-kinase (JAK), son of mother against decapentaplegic (SMAD) and … Product Specific Information This antibody detects endogenous protein at a molecular weight of 42 and 44 kDa. CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 Detection of Human and Mouse Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Western Blot. Tested in Flow Cytometry (Flow) applications. Incubate with rotation for 1–3 hr at 4°C. 770-729-2992 Cart Is Empty Although the identities of all phosphorylation sites in CaD have not been elucidated yet, approximately two-thirds of the total amount of phosphate in CaD … Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde in 1X PBS. Molecular weight of MAP kinase (Erk2) protein is 42 kDa. Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film. A monoclonal antibody (Mab) against uPAR (3B10FC) was generously provided by Dr Robert F Todd III, University of Michigan. The efficacy of these drugs was confirmed by immunoblotting for phospho-ERK1/2 in the case of U0126 and phospho-PKB for LY294002. ERK1/2 are proline-directed kinases that preferentially catalyze the phosphorylation of substrates containing a Pro-Xxx-Ser/Thr-Pro sequence. PBS with 0.02% sodium azide and 50% glycerol pH 7.3. ), please refer to the dye product page for the recommended protocol. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells. Electrotransfer to nitrocellulose membrane (. The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns. Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor, Anti-Rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate). Calculated molecular weight: 38-43 kDa: Observed molecular weight: 38-43 kDa: GenBank accession number: NM_002746: Gene symbol: MAPK3: Gene ID (NCBI) 5595 Conjugate: Unconjugated: Form: Liquid: Purification Method: Antigen affinity purification: Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3. To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold 1X PBS. 80031-1-RR targets Phospho-ERK1/2 (Thr202/Tyr204) in WB,ELISA applications and shows reactivity with Human samples. Nonphosphorylated p44/42 MAPK (Erk1/2) Control Protein: Bacterially expressed, kinase inactive, Erk2 protein serves as a negative control for Western blotting experiments. The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns. ERK1/2 are proline-directed kinases that preferentially catalyze the phosphorylation of substrates containing a Pro-Xxx-Ser/Thr-Pro sequence. Proceed to one of the following specific set of steps. Terminate reaction with 20 µl 3X SDS sample buffer. Human, Mouse, Rat, Hamster, Monkey, Mink, D. melanogaster, Zebrafish, Bovine, Pig, C. elegans. Levels in p-ERK and p-Cdk5 were … CFA-induced p-ERK primarily colocalized with p-Cdk5(S159) in superficial dorsal horn neurons. Western blot analysis of whole cell lysates probed with ERK1/2 antibody followed by detection with HRP conjugated Goat anti Mouse IgG antibody (1/10,000, STAR207P) and visualized on the ChemiDoc™ MP with 36 second exposure. Phospho-p38 MAPK (Thr180/Tyr182) Antibody, Phospho-SAPK/JNK (Thr183/Tyr185) Antibody. ERK1/2 is associated with cell survival, proliferation and development. Scrape cells off the plate and transfer to microcentrifuge tubes. Changing to another country might result in loss of shopping cart. View our alternatives for ab24157 or you can download the archived datasheet PDF from this page. Milli-Mark Anti-Phospho-Erk1/2 Antibody (Thr202/Tyr204, Thr185/Tyr187)-PE This Milli-Mark Phospho-Erk1/2 antibody is validated for use in Flow & WB for the detection of the Milli-Mark Phospho-Erk1/2 protein. Permeabilize cells by adding ice-cold 100% methanol slowly to pre-chilled cells, while gently vortexing, to a final concentration of 90% methanol. europe@ptglab.com, 027-87531629 Transfer supernatant containing phosphorylated substrate to another tube. Resuspend the pellet with 20 µl 3X SDS sample buffer. NOTE: Count cells using a hemocytometer or alternative method. 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. Resuspend cells in approximately 100 µl 4% formaldehyde per 1 million cells. を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために サービス提供条件またはこれに類する合意事項を遵守しなければなりません。. Boster Bio Anti-Phospho-ERK 1/2 (Y222/205) MAPK3 Monoclonal Antibody catalog # M00104Y222. Immunogen: Lane M: Molecular weight ladder. Once activated, Erk1/2 rapidly translocated into the nucleus, where Erk1/2 activates downstream signaling components including Elk-1, although activated Erk1/2 also phosphorylated numerous substrates on (S/T)P sites in all cellular compartments. もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb Copy URL View Product on Manufacturer’s Website Click here to find out more about our Scientific Score! ERK1/2 (phospho-Y204) polyclonal antibody Catalog: AP0490 Host: Rabbit Reactivity: Human,Mouse,Rat BackGround: Store at 4°C short term. NOTE: Please refer to primary antibody product webpage for recommended antibody dilution. Detects a band of approximately 44 kDa (predicted molecular weight: 41 kDa). Anti-phospho-MAPK 1/2 (Thr203/Ty205) Antibody, clone 14B9.1 is a highly specific mouse monoclonal antibody & that targets MAPK/Erk1/2 & has been tested in western blotting. Molecular Biology; Open Reading Frame (ORF) PCR & Amplification ... Anti-phospho-ERK1/2 (pThr 202/Tyr204) antibody produced in rabbit affinity isolated antibody ... Igs) are produced by B lymphocytes and secreted into plasma. CFA-induced p-ERK primarily colocalized with p-Cdk5(S159) in superficial dorsal horn neurons. Resuspend cells in 100 µl of diluted fluorochrome-conjugated secondary antibody (prepared in Antibody Dilution Buffer at the recommended dilution). Wash by centrifugation in Antibody Dilution Buffer or 1X PBS. Keep on ice between washes. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. Aliquot desired number of cells into tubes or wells. The Phospho(Thr202/Tyr204 Thr; 185/Tyr187) /Total ERK1/2 Assay is a sandwich immunoassay (Figure 1) M. SD provdei s a plaet tha thas been pre -coated with capture antibodies for phosphorylated ERK1/2 (Thr202/Tyr204; Thr185/Tyr187) and total ERK1/2on spatially distinct spots . Anti-ERK1 (phospho T202 + Y204) antibody (ab24157) is not available ab24157 is not available and we regret any inconvenience caused. Rabbit polyclonal antibody to ERK1/2 (phospho-Y204) Specificity: p-ERK1/2 (Y204) pAb detects endogenous levels of ERK1 protein when phosphorylated at Tyr204, and ERK2 protein when phosphorylated at Tyr187. Anti-phospho-Erk1/2 (Thr202/Tyr204 Antibody, Thr185/Tyr187), recombinant clone AW39R detects level of phospho-Erk1/2 (Thr202/Tyr204 & has been published & validated for use in Mplex & WB. ASP markedly decreased hepcidin expression by suppressing the expression of JAK1/2, phospho-JAK1/2, phospho-SMAD1/5/8, phospho-ERK1/2, and promoting the expression of SMAD7 in the liver. Arrow points to ERK1/2 (molecular weight 42 & 44 kDa) Inhibition of dynamin II (Dyn II), a GTPase required for endocytosis, with a small molecular weight inhibitor, led to reduced EGF-mediated Stat3 phosphorylation and transcriptional activity in the A431 and HN5 human tumour … Migration of molecular weight markers is indicated. Wash pellet twice with 500 µl 1X kinase buffer. Supplied in SDS Sample buffer. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Here, we examine the role of endocytosis in EGFR signal attenuation and differential signalling. C) Dose-response effect of PGE2 on CSF-1R and ERK1/2 … MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. Anti-phospho-Erk1/2 (Thr202/Tyr204, Thr185/Tyr187)Antibody, recombinant clone AW39R, rabbit monoclonal Anti-phospho-Erk1/2 (Thr202/Tyr204 Antibody, Thr185/Tyr187), recombinant clone AW39R detects level of phospho-Erk1/2 (Thr202/Tyr204 & has been published & validated for use in Mplex & WB. The efficacy of these drugs was confirmed by immunoblotting for phospho-ERK1/2 in the case of U0126 and phospho-PKB for LY294002. Invitrogen Anti-Phospho-ERK1/2 (Thr202, Tyr204) Monoclonal (MILAN8R), eBioscience™, Catalog # 61-9109-42. Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for anti-ERK1/2 Phospho (Thr202/Tyr204) Antibody or a donkey anti-rabbit IgG Antibody conjugated to HRP for anti-GAPDH Antibody (lower). Antibodies to Akt and phospho-Akt (Ser473) were also obtained from Cell Signaling. A number of previous studies suggest that ERK1/2-induced cellular senescence may be associated with the strength of ERK1/2 signals and the duration of its activation (20,77,78,86,90,134). 5595 Vortex, then microcentrifuge for 30 sec at 14,000 x g. Heat the sample to 95–100°C for 2-5 min and microcentrifuge for 1 min at 14,000 x g. Load the sample (15–30 µl) on a 4–20% gel for SDS-PAGE. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration. Several downstream targets of p44/42 have been identified, including p90RSK and the transcription factor Elk-1. Proceed to sample analysis by western immunoblotting or kinase activity (section D). Tested in IHC applications. i got the two bands of 44kDa and 42kDa molecular weight of pERK1 and pERK2 respectively. P27361 Sonicate for 10–15 sec to complete cell lysis and shear DNA (to reduce sample viscosity). Resuspend cells in 200-500 µl of 1X PBS and analyze on flow cytometer. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody. This antibody reacts with Human, Mouse samples. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped … Analyze sample by western blot (see Western Immunoblotting Protocol). This ERK1/2 Antibody is generated from rabbits immunized with a KLH conjugated synthetic phosphopeptide corresponding to amino acid residues surrounding T202/Y204 of human ERK1/2. UniProt ID: いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 Phosphorylated MAP Kinase Control Protein: Fully Rinse three times in 1X PBS for 5 minutes each. NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended. NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase. The phospho-CaD antibody used in this study is specific for the ERK1/2 phosphorylation site at Ser 789 , implicating ERK1/2 in regulation of myometrial contractility. Western blot analysis of ERK1/2 on different lysates. Pellet cells by centrifugation and remove supernatant. Store at –20°C. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. HeLa cells were subjected to SDS PAGE followed by western blot with 80031-1-RR (ERK1/2-phospho-Tyr204 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This antibody reacts with Human, Rat samples. CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST Storage Conditions ERK1/2 functions more frequently as a cellular proliferation marker than as a dual role kinase (37–54). Specificity: Phospho-ERK1/2 (Thr202/Tyr204) Antibody detects endogenous levels of ERK1/2 only when phosphorylated at Threonine 202/Tyrosine 204. いかなる効力も効果も有しません。, 研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 Permeabilize for a minimum of 10 min on ice. Reactivity: Human, Mouse, Rat. Based on these previous studies, cellular proliferation is primarily regulated by the effects of ERK1/2 … They were found during a search for protein kinases that are rapidly phosphorylated after activation of cell surface tyrosine … Fix for 15 min at room temperature (20-25°C). Non-treated PC-3 and Calyculin A treated PC-3 cells were subjected to SDS PAGE followed by western blot with 80031-1-RR Phospho-ERK1/2 (T204) antibody at dilution of 1:5000 incubated at 4℃ overnight. 製品販売条件(CST’s Product Terms of Sale) Incubate for 30 min at room temperature. Inhibition of dynamin II (Dyn II), a GTPase required for endocytosis, with a small molecular weight inhibitor, led to reduced EGF-mediated Stat3 phosphorylation and transcriptional activity in the A431 and HN5 human tumour cell lines. Supplied as 100 µL purified antibody. Isotype controls should be concentration matched and run alongside the primary antibody samples. - Find MSDS or SDS, a COA, data sheets and more information. CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 , P28482, Entrez-Gene Id: Terms and Conditions Anti-phospho-MAPK 1/2 (Thr203/Ty205) Antibody, clone 14B9.1 is a highly specific mouse monoclonal antibody & that targets MAPK/Erk1/2 & has been tested in western blotting. 50ng of Bisphospho-peptide or Non Phosphorylated peptide per dot were adsorbed. Specificity and sensitivity of the Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb: This antibody reacts specifically with as little as 50 pg of phosphorylated MAP kinase and does not cross-react with up to 4 µg of nonphosphorylated MAP kinase by Western blotting. Total cell lysates (15 µg protein) from Jurkat cells without treatment (lane 1) and Jurkat cells starved overnight and then treated with PMA (lane 2) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 0.5 µg/mL Purified anti-ERK1/2 Phospho (Thr202/Tyr204) Antibody (1:1000 dilution), clone 4B11B69 (upper). ERK1 is a 379 amino acid protein with a predicted molecular weight of 43 kD. Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. The size of Erk1/2 is around 45 kDa therefore a 8% PAGE is sufficient to visualize the two bands. (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 min at room temperature. Dot blot analysis of Bi-phospho-ERK1/2-T202/Y204 Antibody (Cat# 102-15492) on nitrocellulose membrane. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when phosphorylated either individually or dually at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2). Many different stimuli, including growth factors, cytokines, virus infection, ligands for heterotrimeric G protein-coupled … ERK1/2 (phospho-Y204) polyclonal antibody Catalog: BS4621 Host: Rabbit Reactivity: Human,Mouse,Rat BackGround: The activation of signal transduction pathways by growth factors, hormones and neurotransmitters is mediated through two closely related MAP kinases, p44 and p42, designated extracellular-signal … Lane M: Molecular weight ladder. For best results examine specimens immediately using appropriate excitation wavelength. CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 Aspirate media from cultures; wash cells with 1X PBS; aspirate. Here, we examine the role of endocytosis in EGFR signal attenuation and differential signalling. Add primary antibody (at the appropriate dilution as recommended in the product datasheet) to 200 µl cell lysate at 1 mg/ml. Anti-phospho-Erk1/2 (Thr202/Tyr204, Thr185/Tyr187)Antibody, recombinant clone AW39R, rabbit monoclonal Anti-phospho-Erk1/2 (Thr202/Tyr204 Antibody, Thr185/Tyr187), recombinant clone AW39R detects level of phospho-Erk1/2 (Thr202/Tyr204 & has been published & validated for use in Mplex & WB. You can still access the archived datasheet PDF below Protect from light. Two similar protein kinases with 85% sequence identity were originally called ERK1 and ERK2. Sonicate on ice three times for 5 sec each. Copyright © 2002-2019 Proteintech Group, Inc. All rights reserved. For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm). Alternatively spliced transcript variants encoding different protein isoforms have been described (provided by RefSeq, Jul 2008). Explore pathways + proteins related to this product. Nonphosphorylated p44/42 MAPK (Erk1/2) Control Protein: Bacterially expressed, kinase inactive, Erk2 protein serves as a negative control for Western blotting experiments. Proteintech The antibody reacts specifically with as little as 0.25 ng of phosphorylated p42 MAP kinase and does not cross-react with up to 4 µg of nonphosphorylated p42 MAP kinase. Guarantee. Discard supernatant. NOTE: Adherent cells or tissue should be dissociated and in single-cell suspension prior to fixation. CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 - Find MSDS or SDS, a COA, data sheets and more information. NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when phosphorylated either individually or dually at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2). Incubate membrane with Anti-rabbit IgG, HRP-linked Antibody (. NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc. The MAPK/ERK cascade plays also a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. Remove PBS and add 0.5 ml ice-cold 1X cell lysis buffer to each plate (10 cm) and incubate on ice for 5 min. および該当する書面のみに従って使用しなければなりません。(e) Incubate specimen in fluorochrome-conjugated secondary antibody diluted in Antibody Dilution Buffer for 1–2 hours at room temperature in dark. From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit. All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow Wash by centrifugation with excess 1X PBS. Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. Aspirate fixative, rinse three times in 1X PBS for 5 minutes each. 7,8 Antibodies to ERK1/2 and phospho-p44/42 ERK1/2 (Thr202/Tyr204) were purchased from Cell Signaling. B)Kineticsof PGE2-induced ERK1/2 phosphorylation. Privacy Policy The Ig molecule in monomeric form is a glycoprotein with a molecular weight of … ERK1/2 (phospho-Y204) polyclonal antibody Catalog: AP0490 Host: Rabbit Reactivity: Human,Mouse,Rat BackGround: Store at 4°C short term. Specifically, we observed biphasic changes in ERK phosphorylation characterized by an increase in phospho-ERK1/2 T202/Y204 at lower compound concentrations and a decrease at higher concentrations . In molecular biology, extracellular signal-regulated kinases or classical MAP kinases are widely expressed protein kinase intracellular signalling molecules that are involved in functions including the regulation of meiosis, mitosis, and postmitotic functions in differentiated cells. Supplied as 100ul in Liquid form antibody. Incubate membrane and primary antibody (at the appropriate dilution and diluent as recommended in the product webpage) in 10 ml primary antibody dilution buffer with gentle agitation overnight at 4°C. Terms of use IMPORTANT: Appropriate isotype controls are highly recommended in order to show specific binding in your primary antibody immunoprecipitation. If the antibody doesn’t perform, you can receive a hassle-free refund or credit note. 07-467 Anti-phospho -MAP Kinase1/2 (Erk1/2) IPFL07810 Immobilon-FL 7 x 8.4 cm PVDF 0.45 mm (sheet) 10/pk 06-642 Anti-phospho -MAP Kinase1/2 (Erk1/2) (Tyr180), polyclonal WBKLS0100 Immobilon Western Chemilu m HRP Substrate 100 mL 17-463 STAR ERK 1/2 ELISA Kit 17-373 Spray & Glow™ ECL WB Detection System 1 ea If necessary, lysate can be stored at -80°C. Storage Conditions Detection of Human and Mouse Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Western Blot. ERK 1, ERK1, ERK1/2, ERT2, HS44KDAP, HUMKER1A, Insulin stimulated MAP2 kinase, MAP kinase 1, MAP kinase 3, MAP kinase isoform p44, MAPK 1, MAPK 3, MAPK3, p44 ERK1, p44 MAPK, P44ERK1, P44MAPK, Phospho-ERK1/2 (Thr202/Tyr204), PRKM3. The membrane was stripped and re-blotted with beta actin antibody as loading control. 1/100 - 1/1000. Mitogen-activated protein kinase 1 (MAPK1) is also known as extracellular signal-regulated kinase 2 (ERK2). The supernatant is the cell lysate. For long term storage, store slides flat at 4°C protected from light. Besides this primary structure requirement, many ERK1/2 substrates possess a D-docking site, an F-docking site, or both. Repeat if necessary. Molecular Weight: ~ 42,44 kDa: ... which was accompanied by sustained increases in phospho-ERK1/2 (p-ERK1/2) and phospho-Cdk5(S159) (p-Cdk5(S159)) in the spinal cord dorsal horn (SCDH). Microcentrifuge for 10 min at 4°C. Western blot analysis of extracts from Hela cells, untreated or treated with TPA (200nM), and NIH/3T3 cells, untreated or treated with PDGF (100ng/ml), using Phospho-Erk1/2 (Thr202/Tyr204) Antibody (upper) or Tubulin (lower). ... Holothurian glycosaminoglycan (hGAG) is a high-molecular-weight form of fucosylated chondroitin sulfate and …