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Androgen receptor Recombinant Rabbit Monoclonal Antibody [PO00-29]

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Specification

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Catalog# HA721156

Androgen receptor Recombinant Rabbit Monoclonal Antibody [PO00-29]

Application

  • IHC-P

  • IF-Tissue

  • IF-Cell

  • IP

  • mIHC

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Androgen receptor Recombinant Rabbit Monoclonal Antibody [PO00-29]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Androgen Receptor aa 300-400 (N terminal).

Species Reactivity

Human, Mouse, Rat

Validated Applications

IHC-P, IF-Tissue, IF-Cell, IP, mIHC, WB

Molecular Weight

Predicted band size: 99 kDa

Positive Control

A549 cell lysate, K-562 cell lysate, 22RV1 cell lysate, rat testis tissue lysates, 22RV1, human prostate carcinoma tissue, human fallopian tube tissue, mouse testis tissue, mouse epididymis tissue, rat epididymis tissue.

Conjugation

unconjugated

Clone Number

PO00-29

RRID

AB_3072278

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-P

  • 1:200-1:1,000

  • WB

  • 1:1,000

  • IF-Tissue

  • 1:50-1:200

  • IF-Cell

  • 1:100

  • IP

  • 1-2μg/sample

  • mIHC

  • 1:3,000

Target

Function

Androgen receptor (AR) is a member of the steroid receptor superfamily that is essential for the growth of prostate cancer cells. As a member of the steroid hormone superfamily, androgen receptor functions within distinct intracellular compartments to mediate cellular growth and transcriptional activation of androgen-regulated signaling pathways. This is accomplished through interactions between steroid ligand and androgen receptor molecules in the cytoplasm that promote dimerization and conformational change. The ligand-receptor homodimers are subsequently translocated to the nucleus for binding to genetic response elements and enhancer regions. Androgen receptor expression is broadly distributed across a variety of normal tissue types as well as in carcinomas of the breast and urothelium, and specifically plays a critical role in prostate development, homeostasis, and carcinogenesis. It has been reported that tyrosine phosphorylation of AR is induced by growth factors and elevated in hormone-refractory prostate tumors. Data suggest that growth factors and their downstream tyrosine kinases, which are elevated during hormone-ablation therapy, can induce tyrosine phosphorylation of AR. Such modification may be important for prostate tumor growth under androgen-depleted conditions. Cellular signaling occurs following androgen binding to the AR and translocation to the nucleus. This activated complex associates with androgen-responsive elements contained in the DNA sequence of target genes, affecting the transcriptional activity of these genes. AR antibody labels epithelial cells and stromal cells in normal prostate. AR reactivity is also found in other types of cells, including epithelial cells of the breast and hepatocytes. In prostate cancer, AR expression is maintained throughout cancer progression. Immunohistochemistry of AR is useful for the evaluation of prostate cancer in routinely processed tissues. The majority of androgen independent hormone refractory prostate cancers express AR. Transcriptional activation of AR is involved in refractory antiandrogen therapy. Androgen receptor expression has been helpful in predicting the clinical response to anti-androgenic treatment.

Background References

1. Aurilio G. et. al. Androgen Receptor Signaling Pathway in Prostate Cancer: From Genetics to Clinical Applications. Cells. 2020 Dec

2. Tesei A. et. al. Editorial: The Androgen Receptor in Breast Cancer. Front Endocrinol (Lausanne). 2021 Jan

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P10275 Human

SwissProt: P19091 Mouse

SwissProt: P15207 Rat

Synonyms

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

Expand

Images

  • Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721156) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human fallopian tube tissue with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human fallopian tube tissue with Rabbit anti-Androgen receptor antibody (HA721156) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721156) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721156) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721156) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat epididymis tissue with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat epididymis tissue with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721156) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Prostate carcinoma<br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/200<br />Antigen retrieval: ER2<br />Platform: Leica Biosystems BOND&reg; RX

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Prostate carcinoma
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/200
    Antigen retrieval: ER2
    Platform: Leica Biosystems BOND® RX

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Mouse<br />Tissue: Epididymis<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>, 1/100, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Mouse
    Tissue: Epididymis
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: HA721156, 1/100, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

  • Immunofluorescence analysis of paraffin-embedded rat epididymis tissue labeling Androgen receptor with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded rat epididymis tissue labeling Androgen receptor with Rabbit anti-Androgen receptor antibody (HA721156) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721156, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Western blot analysis of Androgen receptor on rat testis tissue lysates with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 99 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 3 minutes;<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Androgen receptor on rat testis tissue lysates with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 99 kDa
    Observed band size: 110 kDa

    Exposure time: 3 minutes;

    8% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721156) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Androgen receptor on different lysates with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549 cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: 22RV1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 99 kDa<br />Observed band size: 99/75 kDa<br /><br />Exposure time: 10 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Androgen receptor on different lysates with Rabbit anti-Androgen receptor antibody (HA721156) at 1/1,000 dilution.

    Lane 1: A549 cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: 22RV1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 99 kDa
    Observed band size: 99/75 kDa

    Exposure time: 10 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721156) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of 22RV1 cells labeling Androgen receptor with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Androgen receptor antibody (<a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of 22RV1 cells labeling Androgen receptor with Rabbit anti-Androgen receptor antibody (HA721156) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Androgen receptor antibody (HA721156) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Androgen receptor was immunoprecipitated from 0.2 mg 22RV1 cell lysate with <a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: 22RV1 cell lysate (input)<br />Lane 2: <a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a> IP in 22RV1 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA721156" style="font-weight: bold;text-decoration: underline;">HA721156</a> in 22RV1 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 25 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Androgen receptor was immunoprecipitated from 0.2 mg 22RV1 cell lysate with HA721156 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721156 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: 22RV1 cell lysate (input)
    Lane 2: HA721156 IP in 22RV1 cell lysate
    Lane 3: Rabbit IgG instead of HA721156 in 22RV1 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 25 seconds; ECL: K1801

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Citation