Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187)抗体
参阅全部 Erk1 (pT202/pY204) + Erk2 (pT185/pY187) 一抗
兔多克隆抗体to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)
Rabbit
适用于: WB, IHC-Pmore details
与反应: Mouse, Rat, Human
Synthetic peptide corresponding to Human Erk1 (pT202/pY204) + Erk2 (pT185/pY187). This region is conserved among many species including rat, mouse, cow, frog, snail, nematode, and fruit fly.
(Peptide available as ab5313, ab5354, ab5255)
WB: MDA-MB-231, U-87 MG, Sh-SY5Y, HeLa, PC-12 whole cell lysates, MDA-MB-231 whole cell lysate with treatment of EGF(100 ng/mL for 15 mins. IHC-P: Human breast and colon carcinoma, mouse stomach tissue.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA
BSA is IgG and protease free
Immunogen affinity purified
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the sites of phosphorylation to remove antibody that is reactive with non-phosphorylated ERK 1 + 2. The final product is generated by affinity chromatography using an ERK 1 + 2-derived peptide that is phosphorylated at threonine 202/185 and tyrosine 204/187, respectively, within the activation loop.
多克隆
IgG
Abpromise™承诺保证使用ab4819于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (4) | 1/1000. Predicted molecular weight: 44,42 kDa. |
| IHC-P | 1/10 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue (right) labeling ERK1/2 (pTpY185/187) in the cytoplasm and nucleus with ab4819 at 1/50 dilution, compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab4819 diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819) at 1/1000 dilution
Lane 1 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate, with treatment of EGF(100 ng/mL for 15 mins)
Lane 2 : MDA-MB-231 whole cell lysate
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 4 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 5 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 30 µg per lane.
Predicted band size: 44,42 kDa
Bands of 42 kDa and 44 kDa corresponding to Phospho-p44 MAPK + p42 MAPK pThr185 + pTyr187 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue (right) labeling ERK1/2 (pTpY185/187) in the cytoplasm and nucleus withab4819 at 1/20 dilution, compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab4819 diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue (right) labeling ERK1/2 (pTpY185/187) in the cytoplasm and nucleus with ab4819 at 1/20 dilution, compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab4819 diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819) at 1/1000 dilution
Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate, unstimulated
Lanes 2-6 : PC-12 whole cell lysate, stimulated with 0.5 M sorbitol for 5 minutes
Secondary
All lanes : Goat F (ab')2 anti-rabbit IgG HRP conjugate
Predicted band size: 44,42 kDa
Extracts of PC12 cells were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF.
The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, and then incubated with ab4819 for two hours at room temperature in a 3% BSA-TBST buffer, following its prior incubation with:
Lane 1 and 2: no peptide
Lane 3: the non-phosphopeptide corresponding to the phosphopeptide immunogen
Lane 4: a generic phosphothreonine-containing peptide
Lane 5: a generic phosphotyrosine-containing peptide
Labe 6: the phosphopeptide immunogen
Detection: Pierce SuperSignal™ method.
Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819)
All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody (ab4819) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate, treated with either PDGF
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-rabbit secondary antibody conjugated to Alexa fluor 680
Predicted band size: 44,42 kDa
Data was analyzed on the LI-COR Odyssey® Infrared Imaging System.
