RecombinantRabMAb

Recombinant Anti-Androgen Receptor antibody [EP670Y] (ab52615)

Reviews (4)Q&A (5)References (22)
Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
  • Anti-Androgen Receptor antibody [EP670Y] (ab52615)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP670Y] to Androgen Receptor
  • Suitable for: WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Androgen Receptor antibody [EP670Y]
    See all Androgen Receptor primary antibodies

  • Description

    Rabbit monoclonal [EP670Y] to Androgen Receptor

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Androgen Receptor aa 900 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P10275
    (Peptide available as ab174742)

  • Positive control

    • WB: PC3 whole cell lysate (ab3954), LnCaP and 22Rv1 cell lysate and mouse and rat testis tissue lysates.

  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52615 in the following tested applications.

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

Application Abreviews Notes
WB (2)
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.

We recommend to use 1%SDS Hot lysis prepare method.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.

We recommend to use 1%SDS Hot lysis prepare method.
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

Target

  • Function

    Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins. Transcription activation is down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3.
    Isoform 3 and isoform 4 lack the C-terminal ligand-binding domain and may therefore constitutively activate the transcription of a specific set of genes independently of steroid hormones.

  • Tissue specificity

    Isoform 2 is mainly expressed in heart and skeletal muscle (PubMed:15634333). Isoform 3 is expressed by basal and stromal cells of prostate (at protein level) (PubMed:19244107).

  • Involvement in disease

    Androgen insensitivity syndrome
    Spinal and bulbar muscular atrophy X-linked 1
    Defects in AR may play a role in metastatic prostate cancer. The mutated receptor stimulates prostate growth and metastases development despite of androgen ablation. This treatment can reduce primary and metastatic lesions probably by inducing apoptosis of tumor cells when they express the wild-type receptor.
    Androgen insensitivity, partial

  • Sequence similarities

    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.

  • Domain

    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain. In the presence of bound steroid the ligand-binding domain interacts with the N-terminal modulating domain, and thereby activates AR transcription factor activity. Agonist binding is required for dimerization and binding to target DNA. The transcription factor activity of the complex formed by ligand-activated AR and DNA is modulated by interactions with coactivator and corepressor proteins. Interaction with RANBP9 is mediated by both the N-terminal domain and the DNA-binding domain. Interaction with EFCAB6/DJBP is mediated by the DNA-binding domain.

  • Post-translational
    modifications

    Sumoylated on Lys-388 (major) and Lys-521. Ubiquitinated. Deubiquitinated by USP26. 'Lys-6' and 'Lys-27'-linked polyubiquitination by RNF6 modulates AR transcriptional activity and specificity.
    Phosphorylated in prostate cancer cells in response to several growth factors including EGF. Phosphorylation is induced by c-Src kinase (CSK). Tyr-535 is one of the major phosphorylation sites and an increase in phosphorylation and Src kinase activity is associated with prostate cancer progression. Phosphorylation by TNK2 enhances the DNA-binding and transcriptional activity and may be responsible for androgen-independent progression of prostate cancer. Phosphorylation at Ser-83 by CDK9 regulates AR promoter selectivity and cell growth. Phosphorylation by PAK6 leads to AR-mediated transcription inhibition.
    Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation.

  • Cellular localization

    Nucleus. Cytoplasm. Predominantly cytoplasmic in unligated form but translocates to the nucleus upon ligand-binding. Can also translocate to the nucleus in unligated form in the presence of RACK1.
  • Information by UniProt

  • Database links

    see all

  • Form

    There are 2 isoforms produced by alternative splicing. Isoform 1 is also known as: AR-B; isoform 2 is known as AR-A or variant AR45.

  • Alternative names

    • AIS antibody
    • ANDR_HUMAN antibody
    • Androgen nuclear receptor variant 2 antibody
    • Androgen receptor (dihydrotestosterone receptor; testicular feminization; spinal and bulbar muscular atrophy; Kennedy disease) antibody
    • Androgen receptor antibody
    • androgen receptor splice variant 4b antibody
    • AR antibody
    • AR8 antibody
    • DHTR antibody
    • Dihydro testosterone receptor antibody
    • Dihydrotestosterone receptor (DHTR) antibody
    • Dihydrotestosterone receptor antibody
    • HUMARA antibody
    • HYSP1 antibody
    • KD antibody
    • Kennedy disease (KD) antibody
    • NR3C4 antibody
    • Nuclear receptor subfamily 3 group C member 4 (NR3C4) antibody
    • Nuclear receptor subfamily 3 group C member 4 antibody
    • SBMA antibody
    • SMAX1 antibody
    • Spinal and bulbar muscular atrophy (SBMA) antibody
    • Spinal and bulbar muscular atrophy antibody
    • Testicular Feminization (TFM) antibody
    • TFM antibody
    see all

Images

  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    All lanes : Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/1000 dilution

    Lane 1 : Mouse testis lysate
    Lane 2 : Rat testis lysate
    Lane 3 : Mouse liver lysate
    Lane 4 : Rat liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Diluting buffer: 5% NFDM/TBST

    Loading control: Rabbit monoclonal [EPR16891] to GAPDH (ab181602)

  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    All lanes : Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/1000 dilution

    Lane 1 : LnCAP whole cell lysate (positive control)
    Lane 2 : PC3 whole cell lysate (negative control)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 99 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?
    Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.



    Lanes 1 - 2: Merged signal (red and green). Green - ab52615 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52615 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    All lanes : Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/5000 dilution (Purified)

    Lane 1 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
    Lane 2 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This antibody works better in 1%SDS Hot Lysates in WB. For Lysate preparation protocol, please refer to the protocol here (downloadable copy).

  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/10000 dilution (unpurified) + LnCaP cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 99 kDa
    Observed band size: 99 kDa

  • Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Western blot - Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    All lanes : Anti-Androgen Receptor antibody [EP670Y] (ab52615) at 1/2000 dilution (purified)

    Lane 1 : Mouse testis tissue lysate
    Lane 2 : Rat testis tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-Androgen Receptor antibody [EP670Y] (ab52615)
    Anti-Androgen Receptor antibody [EP670Y] (ab52615)

Protocols

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.

Click here to view the general protocols

References (22)

Publishing research using ab52615? Please let us know so that we can cite the reference in this datasheet.

ab52615 has been referenced in 22 publications.

  • Liva SG  et al. Overcoming resistance to anabolic SARM therapy in experimental cancer cachexia with an HDAC inhibitor. EMBO Mol Med 12:e9910 (2020). PubMed: 31930715
  • Wasmuth EV  et al. Modulation of androgen receptor DNA binding activity through direct interaction with the ETS transcription factor ERG. Proc Natl Acad Sci U S A 117:8584-8592 (2020). PubMed: 32220959
  • Williams ES  et al. Androgen-Dependent Excitability of Mouse Ventral Hippocampal Afferents to Nucleus Accumbens Underlies Sex-Specific Susceptibility to Stress. Biol Psychiatry 87:492-501 (2020). PubMed: 31601425
  • Dauki AM  et al. Transcriptionally Active Androgen Receptor Splice Variants Promote Hepatocellular Carcinoma Progression. Cancer Res 80:561-575 (2020). PubMed: 31685543
  • Acosta-Lopez S  et al. The androgen receptor expression and its activity have different relationships with prognosis in hepatocellular carcinoma. Sci Rep 10:22046 (2020). PubMed: 33328560
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-Androgen Receptor antibody [EP670Y]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEFs)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
MEFs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Mar 05 2021

Western blot abreview for Anti-Androgen Receptor antibody [EP670Y]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (LNCaP cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
25 µg
Specification
LNCaP cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Mar 05 2021

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