Anti-ErbB2 / HER2 抗体 [EP1045Y] (ab134182)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1045Y] to ErbB2 / HER2
- Suitable for: WB, IP, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-ErbB2 / HER2 antibody [EP1045Y]
ErbB2 / HER2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1045Y] to ErbB2 / HER2 -
由来種
Rabbit -
特異性
ab134182 detects ErbB 2 phosphorylated at Tyr1248 as well as unphosphorylated ErbB 2. Mouse species is recommended based on WB results, we do not guarantee IHC-P for mouse.
-
アプリケーション
適用あり: WB, IP, ICC/IF, IHC-Pmore details
適用なし: Flow Cyt -
種交差性
交差種: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- WB: HeLa, SKBR-3, Wild-type HCT 116 and Wild-type A549 cell lysates; IHC-P: Human breast carcinoma tissue; ICC/IF: SKBR cells; IP: HeLa.
-
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
解離定数(KD 値)
KD = 3.00 x 10 -11 M Learn more about KD -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1045Y -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab134182の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
WB |
1/1000. Predicted molecular weight: 137 kDa.
|
|
IP |
1/30.
For unpurifed use at 1/50. |
|
ICC/IF | (2) |
1/250 - 1/500.
|
IHC-P | (1) |
1/1600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified use at 1/100 -1/250. |
特記事項 |
---|
WB
1/1000. Predicted molecular weight: 137 kDa. |
IP
1/30. For unpurifed use at 1/50. |
ICC/IF
1/250 - 1/500. |
IHC-P
1/1600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. For unpurified use at 1/100 -1/250. |
ターゲット情報
-
機能
Protein tyrosine kinase that is part of several cell surface receptor complexes, but that apparently needs a coreceptor for ligand binding. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. GP30 is a potential ligand for this receptor. Regulates outgrowth and stabilization of peripheral microtubules (MTs). Upon ERBB2 activation, the MEMO1-RHOA-DIAPH1 signaling pathway elicits the phosphorylation and thus the inhibition of GSK3B at cell membrane. This prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization.
In the nucleus is involved in transcriptional regulation. Associates with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter and activates its transcription. Implicated in transcriptional activation of CDKN1A; the function involves STAT3 and SRC. Involved in the transcription of rRNA genes by RNA Pol I and enhances protein synthesis and cell growth. -
組織特異性
Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth. -
関連疾患
Hereditary diffuse gastric cancer
Glioma
Ovarian cancer
Lung cancer
Gastric cancer
Chromosomal aberrations involving ERBB2 may be a cause gastric cancer. Deletions within 17q12 region producing fusion transcripts with CDK12, leading to CDK12-ERBB2 fusion leading to truncated CDK12 protein not in-frame with ERBB2. -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
翻訳後修飾
Autophosphorylated. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit (Probable). Ligand-binding increases phosphorylation on tyrosine residues (PubMed:27134172). Signaling via SEMA4C promotes phosphorylation at Tyr-1248 (PubMed:17554007). Dephosphorylated by PTPN12 (PubMed:27134172). -
細胞内局在
Cytoplasm. Nucleus and Cell membrane. Cytoplasm, perinuclear region. Nucleus. Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1. - Information by UniProt
-
参照データベース
- Entrez Gene: 2064 Human
- Entrez Gene: 13866 Mouse
- Omim: 164870 Human
- SwissProt: P04626 Human
- SwissProt: P70424 Mouse
- Unigene: 446352 Human
- Unigene: 290822 Mouse
-
別名
- Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody
- C erb B2/neu protein antibody
- CD340 antibody
see all
画像
-
All lanes : Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/500 dilution
Lane 1 : Wild-type MCF7 cell lysate at 32 µg
Lane 2 : ERBB2 knockout MCF7 cell lysate at 32 µg
Lane 3 : A549 cell lysate at 16 µg
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?Western blot: Anti-ErbB2 / HER2 antibody [EP1045Y] staining at 1/500 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134182 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ERBB2 knockout cell line ab286260 (knockout cell lysate AB300208). To generate this image, wild-type and ERBB2 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature and washed again four times. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. This blot was developed with an ultra high-sensitivity ECL substrate kit and imaged with 20 minutes exposure time.
-
All lanes : Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : ERBB2 knockout A549 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SK-BR-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ErbB2 / HER2 antibody [EP1045Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134182 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type A549 cell lysates with no signal observed at this size in ERBB2 knockout cell line. To generate this image, wild-type and ERBB2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
All lanes : Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/1000 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : ERBB2 knockout HCT 116 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SK-BR-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ErbB2 / HER2 antibody [EP1045Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134182 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in ERBB2 knockout cell line. To generate this image, wild-type and ERBB2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
All lanes : Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ERBB2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 137 kDaFalse colour image of Western blot: Anti-ErbB2 / HER2 antibody [EP1045Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134182 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type HeLa cell lysates with no signal observed at this size in ERBB2 knockout cell line ab255387 (knockout cell lysate ab263758). To generate this image, wild-type and ERBB2 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/1000 dilution + 4T1(Mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling EErbB2 / HER2 with Purified ab134182 at 1:1600 dilution (0.68 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
-
ab134182 (purified) at 1:30 dilution (2µg) immunoprecipitating ErbB2 / HER2 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab134182 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134182 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/10000 dilution (purified) + SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 137 kDa
Blocking and diluting buffer : 5% NFDM/TBST -
Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/1000 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 137 kDa
Blocking and diluting buffer : 5% NFDM/TBST -
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling ErbB2 / HER2 with ab134182 at 1/100 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0).
-
Immunofluorescent analysis of SKBR cells labelling ErbB2 / HER2 with ab134182 at 1/250 dilution.
-
Immunocytochemistry/Immunofluorescence analysis of SK-BR-3 (human mammary gland adenocarcinoma) labelling ErbB2 / HER2 with purified ab134182 at 1/125. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
-
Anti-ErbB2 / HER2 antibody [EP1045Y] (ab134182) at 1/10000 dilution + SKBR-3 cell lysate at 10 µg
Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 137 kDa
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (46)
ab134182 は 46 報の論文で使用されています。
- Li L et al. Neuregulin-1 promotes the proliferation, migration, and angiogenesis of human periodontal ligament stem cells in vitro. Cell Biol Int 46:792-805 (2022). PubMed: 35077607
- Li M et al. IPO7 promotes pancreatic cancer progression via regulating ERBB pathway. Clinics (Sao Paulo) 77:100044 (2022). PubMed: 35588577
- Cheng X et al. Construction and Verification of Immunohistochemistry Parameters-Based Classifier to Predict Local-Recurrence of Upper Tract Urothelial Carcinoma After Kidney-Sparing Surgery. Front Oncol 12:872432 (2022). PubMed: 35600373
- Lu Z et al. Dissecting the genetic and microenvironmental factors of gastric tumorigenesis in mice. Cell Rep 41:111482 (2022). PubMed: 36261019
- Sun C et al. Comparison between core needle biopsy and excisional biopsy for breast neoplasm. Medicine (Baltimore) 100:e26970 (2021). PubMed: 34449464