Anti-SRC3抗体(ab2831)

Submit a review Q&A (1)References (5)
Western blot - Anti-SRC3 antibody (ab2831)
  • Immunocytochemistry/ Immunofluorescence - Anti-SRC3 antibody (ab2831)
  • ChIP - Anti-SRC3 antibody (ab2831)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SRC3 antibody (ab2831)

Key features and details

  • Rabbit polyclonal to SRC3
  • Suitable for: WB, ChIP, IHC-P, ICC/IF
  • Reacts with: Mouse, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-SRC3抗体
    参阅全部 SRC3 一抗

  • 描述

    兔多克隆抗体to SRC3

  • 宿主

    Rabbit

  • 特异性

    Detects amplified in breast cancer 1 (AIB1).

  • 经测试应用

    适用于: WB, ChIP, IHC-P, ICC/IFmore details

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Xenopus laevis

  • 免疫原

    Synthetic peptide corresponding to Human SRC3 aa 3-15.
    Sequence:

    GLGENLDPLASDS


    (Peptide available as ab4915)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 常规说明

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

应用

Our Abpromise guarantee covers the use of ab2831 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 155 kDa. Can be blocked with AIB1 peptide or reuse.
ChIP Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

靶标

  • 功能

    Nuclear receptor coactivator that directly binds nuclear receptors and stimulates the transcriptional activities in a hormone-dependent fashion. Plays a central role in creating a multisubunit coactivator complex, which probably acts via remodeling of chromatin. Involved in the coactivation of different nuclear receptors, such as for steroids (GR and ER), retinoids (RARs and RXRs), thyroid hormone (TRs), vitamin D3 (VDR) and prostanoids (PPARs). Displays histone acetyltransferase activity. Also involved in the coactivation of the NF-kappa-B pathway via its interaction with the NFKB1 subunit. Interacts with PSMB9.

  • 组织特异性

    Widely expressed. High expression in heart, skeletal muscle, pancreas and placenta. Low expression in brain, and very low in lung, liver and kidney.

  • 序列相似性

    Belongs to the SRC/p160 nuclear receptor coactivator family.
    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAS (PER-ARNT-SIM) domain.

  • 结构域

    Contains three Leu-Xaa-Xaa-Leu-Leu (LXXLL) motifs. Motifs 1 and 2 are essential for the association with nuclear receptors, and constitute the RID domain (Receptor-interacting domain).

  • 翻译后修饰

    Acetylated by CREBBP. Acetylation occurs in the RID domain, and disrupts the interaction with nuclear receptors and regulates its function.
    Methylated by CARM1.
    Phosphorylated by IKK complex. Regulated its function.

  • 细胞定位

    Cytoplasm. Nucleus. Mainly cytoplasmic and weakly nuclear. Upon TNF activation and subsequent phosphorylation, it translocates from the cytoplasm to the nucleus.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • ACTR antibody
    • AIB 1 antibody
    • AIB-1 antibody
    • AIB1 antibody
    • Amplified in breast cancer 1 antibody
    • Amplified in breast cancer 1 protein antibody
    • bHLHe42 antibody
    • CAGH 16 antibody
    • CAGH16 antibody
    • CBP interacting protein antibody
    • CBP-interacting protein antibody
    • Class E basic helix-loop-helix protein 42 antibody
    • CTG 26 antibody
    • CTG26 antibody
    • KAT13B / AIB1 antibody
    • MGC141848 antibody
    • NCOA 3 antibody
    • NCoA-3 antibody
    • Ncoa3 antibody
    • NCOA3_HUMAN antibody
    • Nuclear receptor coactivator 3 antibody
    • pCIP antibody
    • RAC 3 antibody
    • RAC-3 antibody
    • RAC3 antibody
    • Receptor associated coactivator 3 antibody
    • Receptor-associated coactivator 3 antibody
    • SRC 3 antibody
    • SRC-3 antibody
    • SRC3 antibody
    • Steroid receptor coactivator protein 3 antibody
    • Thyroid hormone receptor activator molecule 1 antibody
    • TNRC14 antibody
    • TNRC16 antibody
    • TRAM 1 antibody
    • TRAM-1 antibody
    • TRAM1 antibody
    see all

图片

  • Western blot - Anti-SRC3 antibody (ab2831)
    Western blot - Anti-SRC3 antibody (ab2831)
    Western blot of Human 2831 in T47D cell lysate using ab2831.
  • Immunocytochemistry/ Immunofluorescence - Anti-SRC3 antibody (ab2831)
    Immunocytochemistry/ Immunofluorescence - Anti-SRC3 antibody (ab2831)

    ICC/IF image of ab2831 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab2831, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ChIP - Anti-SRC3 antibody (ab2831)
    ChIP - Anti-SRC3 antibody (ab2831)This image is courtesy of Sylvain Daujat

    ab2831 can be used in ChIP. You can expect recruitment of AIB1on the pS2 promoter at some point during transcriptional activation. The amount of DNA precipiated is significant.

    Sonicated Chromatin prepared from untreated or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab2831 to AIB1 and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are % of inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 3 µl of ab2831 and 2x106 cells were used in each ChIP experiment. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SRC3 antibody (ab2831)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SRC3 antibody (ab2831)
    IHC image of ab2831 staining in human melanoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2831, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

文献 (5)

发表研究结果有使用 ab2831？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab2831 被引用在 5 文献中.

  • He Z  et al. SRC3 Is a Cofactor for ROR?t in Th17 Differentiation but Not Thymocyte Development. J Immunol 202:760-769 (2019). PubMed: 30567733
  • You D  et al. Acetylation Enhances the Promoting Role of AIB1 in Breast Cancer Cell Proliferation. Mol Cells 39:663-8 (2016). WB . PubMed: 27665502
  • Mason CE  et al. Location analysis for the estrogen receptor-{alpha} reveals binding to diverse ERE sequences and widespread binding within repetitive DNA elements. Nucleic Acids Res : (2010). ChIP ; Human . PubMed: 20047966
  • Fenne IS  et al. Recruitment of Coactivator GRIP1 to an Estrogen Receptor Transcription Complex is Regulated by the cAMP-dependent Protein Kinase. Endocrinology : (2008). ChIP ; Human . PubMed: 18499756
  • Kawagoe J  et al. Mechanism of the divergent effects of estrogen on the cell proliferation of human umbilical endothelial versus aortic smooth muscle cells. Endocrinology 148:6092-9 (2007). IP, WB ; Human . PubMed: 17872375

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Question

BATCH NUMBER 169016 ORDER NUMBER 134911 DESCRIPTION OF THE PROBLEM No signal or weak signal Only after 1 hour exposition I see a weak band with 80 ?g of a T47D lysate and a little bit stronger band with a 293T extract (transfected cells with human NCoA3). There are some strong unspecific bands at 100 kDa and higher and also at 75 kDa and below SAMPLE I used 293T and T47 cell extracts PRIMARY ANTIBODY Rabbit polycolnal to AIB1 (ab2831) in blocking solution, incuabtion overnight at 4?C dilution 1:500 and 1:000 washing: 3x with TBST DETECTION METHOD ECLplus (Amersham): incubation 5 min POSITIVE AND NEGATIVE CONTROLS USED positive control should be a T47D cell lysate (as described in your data sheet ANTIBODY STORAGE CONDITIONS Aliquots stored at -20?C SAMPLE PREPARATION Buffer: 20 mM Tris/HCl, pH 8; 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 0,2% (v/v) NP-40, 10% Glycerol Protease Inhibitor: Complete EDTA-free (Roche) Phosphataseinhibitors: Phosphatase Inhibitor Cocktail 1 and 2 (Sigma) Denaturation: 5 min 95?C AMOUNT OF PROTEIN LOADED I used 80 ?g of 293T cell lysate and 20, 40 and 80 ?g of T47D cell lysate ELECTROPHORESIS/GEL CONDITIONS reducing conditions, 8% SDS gel. run with 80-120 V TRANSFER AND BLOCKING CONDITIONS transfer with a 3 buffer system, conditons: 1 h, 100 mA (PVDF membrane 9x7 cm) blocking agent: 5% milk powder in TBST (1h, room temparature) SECONDARY ANTIBODY anti rabbit HRP-linked (another company) from donkey, dilution: 1:10000 in blocking solution, incubation: 1h, room temperature washing: 3x with TBST HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I change the dilution from 1:1000 to 1:500 ADDITIONAL NOTES I used the same antibody also in a ChIP experiment and I get also no good results, but in this case it could also be a optimization problem of my ChIP procedure

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Answer

Thank you for the protocol details that you have provided and I'm sorry to hear that you are experiencing difficulty with ab2831. The antibody was characterized for application in Western blotting using T47D cell lysate and a band at 155 kDa was detected. You certainly are loading a sufficient amount of lysate and a suggestion would be to try using nuclear extract instead of whole cell lysate, but the antibody's originator was able to detect AIB1 in the T47D cell lysate. Just to check - is your secondary antibody working properly with other primaries? There have not been any problems with the batch that you received and it sounds to me as if there may be a problem with the particular that you received. I would be happy to send you a free of charge replacement vial or can issue you a credit note or refund. Please let me know how you would like to proceed and I look forward to hearing from you.

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