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Rad51 Recombinant Rabbit Monoclonal Antibody [JM54-26]

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Specification

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Catalog# ET1705-96

Rad51 Recombinant Rabbit Monoclonal Antibody [JM54-26]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

Rad51 Recombinant Rabbit Monoclonal Antibody [JM54-26]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Rad51 aa 1-50 / 339.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 37 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, PANC-1 cell lysate, Raji cell lysate, MEF cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, U-2 OS, NIH/3T3, human ovary cancer tissue, human placenta tissue, human tonsil tissue, mouse spleen tissue, mouse testis tissue, rat spleen tissue, rat testis tissue.

Conjugation

unconjugated

Clone Number

JM54-26

RRID

AB_3070624

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

  • 1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:100

  • IHC-P

  • 1:500-1:1,000

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

Rad51 (RECA, BRCC5) interacts with BRCA1 and BRCA2 to influence subcellular localization and cellular response to DNA damage. BRCA2 inactivation may be a key event leading to genomic instability and tumorigenesis from deregulation of Rad51. Rad52 forms a heptameric ring that binds single-stranded DNA ends and catalyzes DNA-DNA interaction necessary for the annealing of complementary strands. Rad52 can interact with Rad51. Rad54A of the DEAD-like helicase superfamily binds to double-strand DNA and induces a DNA topological change, which is thought to facilitate homologous DNA pairing and stimulate DNA recombination. Rad54B of the DEAD-like helicase superfamily binds to double-stranded DNA and displays ATPase activity in the presence of DNA. Rad54B is abundant in testis and spleen, and mutations of this gene occur in primary lymphoma and colon cancer. MRE11 (meiotic recombination 11, ATLD, HNGS1) is a nuclear 3′-5′ exonuclease/endonuclease that associates with Rad50 and influences homologous recombination, telomere length maintenance, and DNA double-strand break repair. MRE11 is most abundant in proliferating tissues.

Background References

1. Cahyanti D et al. Rad51 Expression in Nasopharyngeal Carcinoma and Its Association with Tumor Reduction: A Preliminary Study in Indonesia. Iran J Pathol 11:155-60 (2016).

2. Arora A et al. Clinicopathological and prognostic significance of RECQL5 helicase expression in breast cancers. Carcinogenesis 37:63-71 (2016).

Sequence Similarity

Belongs to the RecA family. RAD51 subfamily.

Tissue Specificity

Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast.

Post-translational Modification

Ubiquitinated by the SCF(FBH1) E3 ubiquitin ligase complex, regulating RAD51 subcellular location and preventing its association with DNA. Ubiquitinated by RFWD3 in response to DNA damage: ubiquitination leads to degradation by the proteasome, promoting homologous recombination.; Phosphorylated. Phosphorylation of Thr-309 by CHEK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination. Phosphorylation by ABL1 inhibits function.

Subcellular Location

Chromosome, Cytoplasm, Cytoskeleton, Mitochondrion, Nucleus.

UNIPROT

SwissProt: Q06609 Human

SwissProt: Q08297 Mouse

SwissProt: B5DF04 Rat

Synonyms

BRCA1/BRCA2 containing complex, subunit 5 antibody

BRCC 5 antibody

BRCC5 antibody

DNA repair protein RAD51 homolog 1 antibody

DNA repair protein rhp51 antibody

FANCR antibody

hRAD51 antibody

HsRAD51 antibody

HsT16930 antibody

MRMV2 antibody

Expand

Images

  • Western blot analysis of Rad51 on different lysates with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: PANC-1 cell lysate<br />Lane 4: Raji cell lysate<br />Lane 5: MEF cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: RAW264.7 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 37 kDa<br />Observed band size: 37 kDa<br /><br />Exposure time: 3 minutes; 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 Rad51 on different lysates with Rabbit anti-Rad51 antibody (ET1705-96) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: PANC-1 cell lysate
    Lane 4: Raji cell lysate
    Lane 5: MEF cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: RAW264.7 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 37 kDa
    Observed band size: 37 kDa

    Exposure time: 3 minutes; 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 (ET1705-96) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 U-2 OS cells labeling Rad51 with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol 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-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</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 U-2 OS cells labeling Rad51 with Rabbit anti-Rad51 antibody (ET1705-96) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol 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-Rad51 antibody (ET1705-96) 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Rad51 with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/200 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-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/200 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 NIH/3T3 cells labeling Rad51 with Rabbit anti-Rad51 antibody (ET1705-96) at 1/200 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-Rad51 antibody (ET1705-96) at 1/200 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 ovary cancer tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</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 ovary cancer tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) 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 placenta tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</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 placenta tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) 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 tonsil tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 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 tonsil tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) at 1/500 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 spleen tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 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 spleen tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) at 1/500 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-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</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-Rad51 antibody (ET1705-96) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) 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 spleen tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 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 spleen tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) at 1/500 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 testis tissue with Rabbit anti-Rad51 antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>) at 1/500 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 testis tissue with Rabbit anti-Rad51 antibody (ET1705-96) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-96) at 1/500 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 U-2 OS cells labeling Rad51.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1705-96" style="font-weight: bold;text-decoration: underline;">ET1705-96</a>, 1/1,000) (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 U-2 OS cells labeling Rad51.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-96, 1/1,000) (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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Citation