Anti-Glucocorticoid Receptor alpha 抗体 - ChIP Grade (ab3580)

製品の概要

  • 製品名Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade
  • 製品の詳細
    Rabbit polyclonal to Glucocorticoid Receptor alpha - ChIP Grade
  • アプリケーション適用あり: IHC-P, Flow Cyt, ICC, IP, WB, ChIP, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Sheep, Dog, Human, Non Human Primates, Plants
    交差が予測される動物種: Rabbit
  • 免疫原

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

    CEIITNQIPKYSNGNIKK


    (Peptide available as ab39764)

  • 特記事項

    GR alpha proteins has many isoforms e.g. GR alpha-A, Alpha-2, GR-A alpha, Alpha-B so multiple bands can be observed in WB. Please check Uniprot database or PMID 15866175 for more information.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3580 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P 1/20.
EMSA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
ICC Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 4 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 86 kDa).Can be blocked with Human Glucocorticoid Receptor alpha peptide (ab39764).
ChIP Use at an assay dependent concentration.
ICC/IF 1/100 - 1/200.

ターゲット情報

  • 関連性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.
  • 細胞内局在Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand; nuclear after ligand-binding.
  • 参照データベース
  • 別名
    • 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

Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade 画像

  • Immunocytochemistry/Immunofluorescence analysis of U251 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa 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.

  • Immunocytochemistry/Immunofluorescence analysis of A2058 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.

  • ab3580 staining Glucocorticoid Receptor alpha in 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.

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  • ICC/IF image of ab3580 stained Hek293 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3580, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • ab3580 staining glucocorticoid receptor in serum starved HeLa 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.

  • 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.
  • 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.

  • 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.

  • 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.

Anti-Glucocorticoid Receptor alpha antibody - ChIP Grade (ab3580) 使用論文

This product has been referenced in:
  • Zinkhan EK  et al. Maternal tobacco smoke increased visceral adiposity and serum corticosterone levels in adult male rat offspring. Pediatr Res 76:17-23 (2014). WB ; Rat . Read more (PubMed: 24727947) »
  • Mparmpakas D  et al. Differential expression of placental glucocorticoid receptors and growth arrest-specific transcript 5 in term and preterm pregnancies: evidence for involvement of maternal stress. Obstet Gynecol Int 2014:239278 (2014). WB ; Human . Read more (PubMed: 24899900) »

See all 8 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

投稿 Feb 14 2014

Abreviews
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
Username

Mr. Chris Zelinka

Verified customer

投稿 Jun 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 May 03 2013

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 ...

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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 ...

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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 cor...

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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 a...

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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 ...

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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...

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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 ...

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