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Epigenetics and Nuclear Signaling Transcription Domain Families Zinc Finger

Anti-Glucocorticoid Receptor alpha抗体(ab3580)

  • Datasheet
Reviews (3)Q&A (10)References (26)

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Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
  • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
  • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
  • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

Key features and details

  • Rabbit polyclonal to Glucocorticoid Receptor alpha
  • Suitable for: IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-Glucocorticoid Receptor alpha抗体
  • 描述

    兔多克隆抗体to Glucocorticoid Receptor alpha
  • 宿主

    Rabbit
  • 经测试应用

    适用于: IHC-P, ICC/IFmore details
  • 种属反应性

    与反应: Human
  • 免疫原

    Synthetic peptide corresponding to Human Glucocorticoid Receptor alpha aa 755-771.
    Sequence:

    CEIITNQIPKYSNGNIKK


    Database link: P04150
    (Peptide available as ab39764)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • IHC: human cervical carcinoma, heart tissue, tonsil tissue; ICC: U251, A2058, HeLa
  • 常规说明

    GR alpha proteins has many isoforms e.g. GR alpha-A, Alpha-2, GR-A alpha, Alpha-B. Please check Uniprot database or PMID 15866175 for more information.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

性能

  • 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 or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.05% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Zinc Finger
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Corticoid
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Corticoid
    • Neuroscience
    • Processes

相关产品

  • Assay kits

    • Glucocorticoid Receptor Transcription Factor Assay Kit (Colorimetric) (ab207207)
  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • Human Glucocorticoid Receptor alpha peptide (ab39764)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Rosiglitazone, PPARgamma agonist (ab120762)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3580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
IHC-P (1)
1/20.
ICC/IF (1)
1/100 - 1/200.
说明
IHC-P
1/20.
ICC/IF
1/100 - 1/200.

靶标

  • 相关性

    Glucocorticoids are a family of steroids necessary for the regulation of energy metabolism and the immune and inflammatory responses. These compounds exert their effect through their interaction with the Glucocorticoid Receptor (GR) and that complex's subsequent association with DNA. All normal mammalian tissues examined to date have been shown to contain GR. The human GR exists in two forms, alpha and beta, which are thought to be the result of alternative splicing of a single gene. Sequence analysis indicates that alpha and beta forms of human GR are 777 and 742 amino acids long, respectively. They are identical up to residue 727, after which they diverge. After ligand binding, the 94 kDa GR alpha isoform translocates from the cytoplasm to the nucleus where it regulates gene expression. In contrast, the 90 kDa GR beta isoform does not appear to bind either glucocorticoid agonists or antagonists, and has been localized predominantly in the nucleus independent of hormone treatment in some human cell lines. Studies suggest that human GR alpha has a greater affinity for GR response elements (GREs) than GR beta only when in the ligand bound state.
  • 细胞定位

    Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand; nuclear after ligand-binding.
  • 数据库链接

    • Entrez Gene: 2908 Human
    • SwissProt: P04150 Human
    • 别名

      • GCCR antibody
      • GCR antibody
      • Glucocorticoid receptor alpha isoform antibody
      • Glucocorticoid receptor antibody
      • GR antibody
      • GRL antibody
      • NR3C1 antibody
      • Nuclear receptor subfamily 3 group C member 1 antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

      Immunocytochemistry/Immunofluorescence analysis of U-251 MG (Human brain glioma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)

      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial adenocarcinoma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)

      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human cervical carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

      Immunocytochemistry/Immunofluorescence analysis of A2058 (Human metastatic melanoma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/200. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody (ab3580)

      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human heart tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

      ab3580 staining glucocorticoid receptor in serum starved HeLa (Human epithelial adenocarcinoma cell line) cells treated with rosiglitazone (ab120762), by ICC/IF. Changes in localization of glucocorticoid receptor (translocation from cytoplasm to nucleous) correlates with increased concentration of rosiglitazone, as described in literature.
      The cells were incubated at 37°C for 1h in media containing different concentrations of ab120762 (rosiglitazone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab3580 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)This image is courtesy of an anonymous Abreview

      ab3580 staining Glucocorticoid Receptor alpha in mouse epididymis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with Bouin's solution and blocked with 1.5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in blocking buffer) for 14 hours at 4°C. ab6721 Goat anti-rabbit HRP (1/200) was used as the secondary antibody.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580)

      ab3580 (1µg/ml) staining glucocorticoid receptor alpha in human hippocampus using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic staining in the neuropil and blood vessel smooth muscle.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

    实验方案

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet download

      Download

    文献 (26)

    发表研究结果有使用 ab3580?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3580 被引用在 26 文献中.

    • Zhou Z  et al. LncRNA XIST promotes inflammation by downregulating GRa expression in the adenoids of children with OSAHS. Exp Ther Med 21:500 (2021). PubMed: 33791009
    • Vega N  et al. Exposure to pollutants altered glucocorticoid signaling and clock gene expression in female mice. Evidence of tissue- and sex-specificity. Chemosphere 262:127841 (2021). PubMed: 32784060
    • Wang SB  et al. Increased lipopolysaccharide content is positively correlated with glucocorticoid receptor-beta expression in chronic rhinosinusitis with nasal polyps. Immun Inflamm Dis 8:605-614 (2020). PubMed: 32870597
    • Ariel F  et al. R-Loop Mediated trans Action of the APOLO Long Noncoding RNA. Mol Cell 77:1055-1065.e4 (2020). PubMed: 31952990
    • Xu W  et al. Huai Qi Huang Potentiates Dexamethasone-Mediated Lethality in Acute Lymphoblastic Leukemia Cells by Upregulating Glucocorticoid Receptor a. Med Sci Monit 26:e921649 (2020). PubMed: 32065117
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    32 µg
    Gel Running Conditions
    Reduced Denaturing (10% gel, semi-dry transfer)
    Sample
    Chicken Cell lysate - whole cell (LMH overexpressing chicken GR)
    Specification
    LMH overexpressing chicken GR
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Feb 14 2014

    Immunocytochemistry/ Immunofluorescence

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 20 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Sample
    Chicken Cell (retina)
    Specification
    retina
    Permeabilization
    Yes - triton X-100 in PBS
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Chris Zelinka

    Verified customer

    提交于 Jun 04 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (epididymis)
    Specification
    epididymis
    Fixative
    bouin's solution
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 May 03 2013

    Question

    Hello,

    We have a quality issue for item ab3580. Please check attached file for detail.

    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 29 2013

    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

    I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Abcam Scientific Support

    回复于 Jan 29 2013

    Question

    High background and no specific results using ab3580 in Western blot and ChIP.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 30 2012

    Answer

    Thank you very much for your call today and for letting us know about the trouble with this antibody.

    As we discussed, please forward me anyimages showing the results with this antibody, and I will get in touch with the lab.

    I look forward to hearing from you. I apologize for the inconvenience, and if there is anything else that we can do for you please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    回复于 Jul 30 2012

    Question

    Phone call reporting problems using ab3580 in WB.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 06 2012

    Answer

    Thank you for contacting us and reporting the problems you have been experiencing with anti-Glucocorticoid Receptor alpha antibody (ab3580). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As discussed over the phone, I have checked into how this antibody has been characterised by Western blotting. This antibody has not been characterised in-house by Western blotting but we have had reports of it being used for this application from journals published (please see the attached journal articles).


    If you wouldn't mind, could you fill out thequestionnaire I have attached to this email so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results with ab3580. This information will also allow us to investigate this case internally and initiate any additional testing where necessary. If you could include an image of the results you have obtained that would be most useful.


    If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody. In order to do this we need the order number (or the delivery address and the approximate date the order was placed).

    I look forward to receiving your reply.

    Read More

    Abcam Scientific Support

    回复于 Feb 06 2012

    Question

    Are any of your glucocorticoid receptor antibodies specific for the GR-alpha isoform? Is the predicted seequence of GR-beta short enough to be visibly different on a standard western blot?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 02 2007

    Answer

    Thank you for your interests in our products. Yes, we have a product (ab3580) that specifically detects GR-alpha from human, mouse and rat tissues and does not detect GR-beta isoform. I have attached a datasheet link to this product for your easy access. As you know, the human GR exists in two forms, alpha and beta, which are thought to be the result of alternative splicing of a single gene. Sequence analysis indicates that the alpha and beta forms of human GR are 777 and 742 amino acids long, respectively. They are identical up to residue 727, after which they diverge. After ligand binding, the 94 kDa GR alpha isoform has been observed to translocate from the cytoplasm to the nucleus where it regulates gene expression. In contrast, the 90 kDa GR beta isoform does not appear to bind either glucocorticoid agonists or antagonists, and has been localized predominantly in the nucleus independent of hormone treatment in some human cell lines. Studies suggest that human GR beta might function as a dominant negative inhibitor of GR alpha activity.

    Read More

    Abcam Scientific Support

    回复于 Apr 02 2007

    Question

    I got a further question on the ChIP-grade GR antibody. You say that this antibody is guaranteed to work in ChIP. But I've had problem previously with the sonication and immunoprecipitation steps of the ChIP experiments. I've noticed that I need SDS (at least 0.5%) in the sonication buffer to get efficient sonication of my chromatin (using the bioruptor sonicator). This high detergent concentration sometimes interferes with the immunoprecipitation step, as some antibodies don't do well in such conditions. I've noticed that your published ChIP protocol does not include any SDS in the sonication buffer, which would result in incomplete fragmentation of my chromatin using the bioruptor. I am worried that adding SDS might affect the immunoprecipitation step. So, basically my question is in what ChIP conditions was this antibody tested? You say that this antibody (ab3580) was shown to be ChIP-grade by an external originator... but was protocol/conditions was he using? The published AbCam one or something else?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 13 2006

    Answer

    Thank you for getting back in touch with me. Yes, you are correct a high SDS content can potentially interfere with your antibody-antigen interaction by ChIP. However, our Abcam protocol has been optimized and developed by an experienced chromatin postdoc colleague in our lab. She also uses the bioruptor for the sonication step. With regards this antibody; I can tell you that Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580) was actually applied using ChIP in the following publication: Ichijo T et al. The Smad6-histone deacetylase 3 complex silences the transcriptional activity of the glucocorticoid receptor: potential clinical implications. J Biol Chem 280:42067-77 (2005). PubMed: 16249187 I have emailed this publication in a follow up email for your information. You may wish to consult the chromatin IP methods employed.

    Read More

    Abcam Scientific Support

    回复于 Jul 17 2006

    Question

    I am interested in testing both ab3578 and ab3580 in a fish cell culture system, trout and sea bream hepatocyte cultures. There is no technical data available considering testing in these species. My question is economic. Is it possible to acquire smaller aliquot sizes in order to test the antibodies by western blot in these species. As no cross-reactivity will result in an expensive exercise >600E. Does this possibility exist at Abcam?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 16 2005

    Answer

    Thank you for your enquiry. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in species or applications for which we do not have any information, please let us know how you get on and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on a number of rewards.

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    Abcam Scientific Support

    回复于 Mar 16 2005

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Strong band between 66 - 98 kDa markers, therefore it is not the 94 kDa that would be present according to the datasheet. SAMPLE HUVEC whole cell lysate PRIMARY ANTIBODY ab-3580 at 4 microgrammes/mL in 5% milk/TBST SECONDARY ANTIBODY Anti-rabbit HRP (Amersham) 1:5000 in 5% milk/TBST DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED We would like you to send us a positive control (either recombinant human GR alpha or HeLa cell lysates, which I believe are used as positive controls for GR alpha and COS-74 cell lysates, which would be a negative control since they are strongly positive for the beta isoform and not alpha) for this antibody since it is not performing as anticipated from the datasheet. SAMPLE PREPARATION Boiled for 3 min in sample buffer (beta-mercapto, SDS, bromophenol blue etc.) AMOUNT OF PROTEIN LOADED 20 microgrammes ELECTROPHORESIS/GEL CONDITIONS 7.5 % Tris-Glycine reducing gel run for 1 h at TRANSFER AND BLOCKING CONDITIONS Transferred at 100 V for 1.5 h (Trizma/Glycine transfer buffer) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? once HAVE YOU RUN A "NO PRIMARY" CONTROL? No

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    Abcam community

    Verified customer

    Asked on Feb 02 2005

    Answer

    Thank you for the details that you have provided and I'm sorry to hear that you are experiencing difficulty with this antibody. As stated on the online datasheet, we do recommend using HeLa cell lysate as a positive control for ab3580. Without running a positive control, it is difficult to say what may be going on. However, I unfortunately can't send you the lysate free of charge. If you have any additional questions, please let us know.

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    Abcam Scientific Support

    回复于 Feb 02 2005

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