重组Anti-ErbB2 / HER2抗体[EP1045Y] - BSA and Azide free (ab194979)
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 - BSA and Azide free
- Suitable for: WB, ICC/IF, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-ErbB2 / HER2抗体[EP1045Y] - BSA and Azide free
参阅全部 ErbB2 / HER2 一抗 -
描述
兔单克隆抗体[EP1045Y] to ErbB2 / HER2 - BSA and Azide free -
宿主
Rabbit -
特异性
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.
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经测试应用
适用于: WB, ICC/IF, IHC-P, IPmore details
不适用于: Flow Cyt -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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.
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常规说明
ab194979 is the carrier-free version of ab134182.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
解离常数(KD)
KD = 3.00 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1045Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab194979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 |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 185 kDa (predicted molecular weight: 137 kDa).
Please check the parent abID, ab134182, for more information on dilutions. |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 185 kDa (predicted molecular weight: 137 kDa). Please check the parent abID, ab134182, for more information on dilutions. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
靶标
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功能
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. -
组织特异性
Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth. -
疾病相关
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. -
序列相似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
翻译后修饰
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). -
细胞定位
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
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数据库链接
- Entrez Gene: 2064 Human
- Entrez Gene: 13866 Mouse
- Omim: 164870 Human
- SwissProt: P04626 Human
- SwissProt: P70424 Mouse
- Unigene: 446352 Human
- Unigene: 290822 Mouse
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别名
- Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody
- C erb B2/neu protein antibody
- CD340 antibody
see all
图片
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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?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling ErbB2 / 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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Anti-ErbB2 / HER2 antibody [EP1045Y] - BSA and Azide free (ab194979) + SKBR-3 (human mammary gland adenocarcinoma) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 137 kDa
Observed band size: 185 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
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Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling ErbB2 / HER2 with ab134182 at 1/100 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134182).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (7)
ab194979 被引用在 7 文献中.
- Xie H & Wang H PRL-3 promotes breast cancer progression by downregulating p14ARF-mediated p53 expression. Oncol Lett 15:2795-2800 (2018). PubMed: 29435006
- Chan KK et al. Targeting estrogen receptor subtypes (ERa and ERß) with selective ER modulators in ovarian cancer. J Endocrinol 221:325-36 (2014). Human . PubMed: 24819599
- Sollome JJ et al. HER2/HER3 regulates extracellular acidification and cell migration through MTK1 (MEKK4). Cell Signal 26:70-82 (2013). Human . PubMed: 24036211
- Zhang YJ et al. pH-responsive artemisinin derivatives and lipid nanoparticle formulations inhibit growth of breast cancer cells in vitro and induce down-regulation of HER family members. PLoS One 8:e59086 (2013). WB ; Human . PubMed: 23516601
- Ariazi EA et al. Estrogen induces apoptosis in estrogen deprivation-resistant breast cancer through stress responses as identified by global gene expression across time. Proc Natl Acad Sci U S A 108:18879-86 (2011). WB ; Human . PubMed: 22011582
- Behnsawy HM et al. Expression of cell cycle-associated proteins in non-muscle-invasive bladder cancer: correlation with intravesical recurrence following transurethral resection. Urol Oncol 29:495-501 (2011). IHC-P ; Human . PubMed: 19914103
- Kawano S et al. Silencing of ErbB3/ErbB2 signaling by immunoglobulin-like Necl-2. J Biol Chem 284:23793-805 (2009). PubMed: 19561085