重组Anti-STAT5a抗体[E289] (ab32043)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E289] to STAT5a
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-STAT5a抗体[E289]
参阅全部 STAT5a 一抗 -
描述
兔单克隆抗体[E289] to STAT5a -
宿主
Rabbit -
特异性
The antibody recognises Stat5a. It does not cross-react with other Stat family members. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. -
经测试应用
适用于: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, 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: A431 cell lysate. IHC-P: Human squamous lung carcinoma. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells.
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常规说明
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E289 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32043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 | (1) |
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa).
|
IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
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ICC/IF |
1/100 - 1/500.
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IP |
1/20.
For unpurified use at 1/80. |
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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.For unpurified use at 1/10. |
说明 |
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WB
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
ICC/IF
1/100 - 1/500. |
IP
1/20. For unpurified use at 1/80. |
Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.For unpurified use at 1/10. |
靶标
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功能
Carries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription. -
序列相似性
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
翻译后修饰
Tyrosine phosphorylated in response to IL-2, IL-3, IL-7, IL-15, GM-CSF, growth hormone, prolactin, erythropoietin and thrombopoietin. Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity. -
细胞定位
Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. - Information by UniProt
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数据库链接
- Entrez Gene: 6776 Human
- Entrez Gene: 20850 Mouse
- Entrez Gene: 24918 Rat
- Omim: 601511 Human
- SwissProt: P42229 Human
- SwissProt: P42230 Mouse
- SwissProt: Q62771 Rat
- Unigene: 437058 Human
see all -
别名
- Mammary gland factor antibody
- MGF antibody
- Signal transducer and activator of transcription 5A antibody
see all
图片
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Anti-STAT5a antibody [E289] (ab32043) at 1/20000 dilution (purified) + K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling STAT5a with purified ab32043 at 1:1000 dilution (0.12 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use). PBS instead of the primary antibody was used as the negative control.
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Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling STAT5a with purified ab32043 at 1:100 (1.2 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling STAT5a with purified ab32043 at 1/100 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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ab32043 (purified) at 1:20 dilution (0.6ug) immunoprecipitating in TF-1 whole cell lysate. TF-1 (Human Erythroleukemia erythroblast) whole cell lysate 10ug
Lane 2 (+): ab32043 & TF-1 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32043 in TF-1 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
Unpurified ab32043 at 1/250 dilution, staining human squamous lung carcinoma by Immunohistochemistry, paraffin-embedded tissue. Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (Purified)
Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling STAT5a with unpurified ab32043 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control: PBS only.
Nuclear counter stain: DAPI. -
Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (unpurified) + A431 cell lysate
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (unpurified) +
Recombinant Human STAT5a protein (ab84627) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 91 kDa
Exposure time: 20 seconds -
Overlay histogram showing Jurkat cells stained with unpurified ab32043 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32043, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
Tissue Microarrays stained for "Anti-STAT5a antibody [E289]” using "ab32043" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab32043 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (38)
ab32043 被引用在 38 文献中.
- Yang Y et al. Targeting the STAT5A/IDO1 axis overcomes radioresistance and reverses the immunosuppressive tumor microenvironment in NSCLC. Int J Oncol 62:N/A (2023). PubMed: 36453241
- Able AA et al. TNFα Effects on Adipocytes Are Influenced by the Presence of Lysine Methyltransferases, G9a (EHMT2) and GLP (EHMT1). Biology (Basel) 12:N/A (2023). PubMed: 37237488
- Sorger H et al. Blocking STAT3/5 through direct or upstream kinase targeting in leukemic cutaneous T-cell lymphoma. EMBO Mol Med 14:e15200 (2022). PubMed: 36341492
- Venigalla S et al. MED12 Regulates Human Adipose-Derived Stem Cell Adipogenesis and Mediator Kinase Subunit Expression in Murine Adipose Depots. Stem Cells Dev 31:119-131 (2022). PubMed: 35018809
- Chen C et al. A highly selective JAK3 inhibitor is developed for treating rheumatoid arthritis by suppressing γc cytokine-related JAK-STAT signal. Sci Adv 8:eabo4363 (2022). PubMed: 35984890