Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) 抗体 (ab4819)
Key features and details
- Rabbit polyclonal to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)
- Suitable for: WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
製品の概要
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製品名
Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody
Erk1 (pT202/pY204) + Erk2 (pT185/pY187) 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Erk1 (pT202/pY204) + Erk2 (pT185/pY187) -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details -
種交差性
交差種: Mouse, Human -
免疫原
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ポジティブ･コントロール
- 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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
バッファー
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
精製度
Immunogen affinity purified -
特記事項（精製）
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. -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
アプリケーション
Our Abpromise guarantee covers the use of ab4819 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB | 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. |
画像
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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.
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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 kDaBands 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.
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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.
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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.
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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 kDaExtracts 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.
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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 kDaData was analyzed on the LI-COR Odyssey® Infrared Imaging System.
プロトコール
データシートおよび資料
参考文献 (23)
ab4819 は 23 報の論文で使用されています。
- Li W et al. Berberine suppresses IL-33-induced inflammatory responses in mast cells by inactivating NF-?B and p38 signaling. Int Immunopharmacol 66:82-90 (2019). PubMed: 30445310
- Ursoli Ferreira F et al. Endothelial Cells Tissue-Specific Origins Affects Their Responsiveness to TGF-ß2 during Endothelial-to-Mesenchymal Transition. Int J Mol Sci 20:N/A (2019). PubMed: 30678183
- Fa Z et al. MicroRNA-150 suppresses the growth and malignant behavior of papillary thyroid carcinoma cells via downregulation of MUC4. Exp Ther Med 16:45-52 (2018). PubMed: 29896226
- Kole D et al. High molecular weight FGF2 isoforms demonstrate canonical receptor-mediated activity and support human embryonic stem cell self-renewal. Stem Cell Res 21:106-116 (2017). PubMed: 28433654
- Hao GM et al. The Hippo signaling pathway: a potential therapeutic target is reversed by a Chinese patent drug in rats with diabetic retinopathy. BMC Complement Altern Med 17:187 (2017). PubMed: 28372586
