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hHR23A Recombinant Rabbit Monoclonal Antibody [JE38-49]

Usd: 385

Specification

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Catalog# HA722215

hHR23A Recombinant Rabbit Monoclonal Antibody [JE38-49]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

Conjugation

  • unconjugated

Overview

Product Name

hHR23A Recombinant Rabbit Monoclonal Antibody [JE38-49]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human hHR23A aa 181-280 / 363.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 40 kDa

Positive Control

SK-OV-3 cell lysate, RPMI 8226 cell lysate, 786-0 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, COS-1 cell lysate, SK-OV-3, human liver tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

JE38-49

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:1,000

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

Target

Function

UV excision repair protein RAD23 homolog A is a protein that in humans is encoded by the RAD23A gene. The protein encoded by this gene is one of two human homologs of Saccharomyces cerevisiae Rad23, a protein involved in nucleotide excision repair (NER). This protein was shown to interact with, and elevate the nucleotide excision activity of 3-methyladenine-DNA glycosylase (MPG), which suggested a role in DNA damage recognition in base excision repair. This protein contains an N-terminal ubiquitin-like domain, which was reported to interact with 26S proteasome, as well as with ubiquitin protein ligase E6AP, and thus suggests that this protein may be involved in the ubiquitin mediated proteolytic pathway in cells.

Background References

1. Byeon IL et al. Structure of HIV-1 Vpr in complex with the human nucleotide excision repair protein hHR23A. Nat Commun. 2021 Nov

2. Becht DC et al. Residual Structure in the Denatured State of the Fast-Folding UBA(1) Domain from the Human DNA Excision Repair Protein HHR23A. Biochemistry. 2022 May

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P54725 Human

SwissProt: P54726 Mouse

Synonyms

hHR 23A antibody

hHR23A antibody

HR23A antibody

MGC111083 antibody

RAD 23a antibody

RAD23 homolog A (S. cerevisiae) antibody

RAD23 homolog A antibody

RAD23 yeast homolog A antibody

RAD23A antibody

RD23A_HUMAN antibody

Expand

Images

  • Western blot analysis of hHR23A on different lysates with Rabbit anti-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</a>) at 1/2,000 dilution.<br /><br />Lane 1: SK-OV-3 cell lysate<br />Lane 2: RPMI 8226 cell lysate<br />Lane 3: 786-0 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 40 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<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/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</a>) at 1/2,000 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 hHR23A on different lysates with Rabbit anti-hHR23A antibody (HA722215) at 1/2,000 dilution.

    Lane 1: SK-OV-3 cell lysate
    Lane 2: RPMI 8226 cell lysate
    Lane 3: 786-0 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 40 kDa
    Observed band size: 52 kDa

    Exposure time: 20 seconds; ECL: K1801;

    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 (HA722215) at 1/2,000 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 SK-OV-3 cells labeling hHR23A with Rabbit anti-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</a>) at 1/1,000 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-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</a>) at 1/1,000 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 SK-OV-3 cells labeling hHR23A with Rabbit anti-hHR23A antibody (HA722215) at 1/1,000 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-hHR23A antibody (HA722215) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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 liver tissue with Rabbit anti-hHR23A antibody (HA722215) 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 (HA722215) 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 liver tissue with Rabbit anti-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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 mouse liver tissue with Rabbit anti-hHR23A antibody (HA722215) 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 (HA722215) 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 rat liver tissue with Rabbit anti-hHR23A antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</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 rat liver tissue with Rabbit anti-hHR23A antibody (HA722215) 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 (HA722215) 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.

  • Flow cytometric analysis of SK-OV-3 cells labeling hHR23A.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722215" style="font-weight: bold;text-decoration: underline;">HA722215</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SK-OV-3 cells labeling hHR23A.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722215, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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