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RAP80 Mouse Monoclonal Antibody [A1B11]

Usd: 350

Specification

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Catalog# EM1902-05

RAP80 Mouse Monoclonal Antibody [A1B11]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

RAP80 Mouse Monoclonal Antibody [A1B11]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human RAP80 aa 330-524 / 719.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Molecular Weight

80 kDa

Positive Control

Siha cell lysates, rat testis tissue, mouse testis tissue, SiHa cell.

Conjugation

unconjugated

Clone Number

A1B11

RRID

AB_3068949

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

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

Ubiquitin-binding protein. Specifically recognizes and binds 'Lys-63'-linked ubiquitin. Plays a central role in the BRCA1-A complex by specifically binding 'Lys-63'-linked ubiquitinated histones H2A and H2AX at DNA lesions sites, leading to target the BRCA1-BARD1 heterodimer to sites of DNA damage at double-strand breaks (DSBs). The BRCA1-A complex also possesses deubiquitinase activity that specifically removes 'Lys-63'-linked ubiquitin on histones H2A and H2AX. Also weakly binds monoubiquitin but with much less affinity than 'Lys-63'-linked ubiquitin. May interact with monoubiquitinated histones H2A and H2B; the relevance of such results is however unclear in vivo. Does not bind Lys-48'-linked ubiquitin. May indirectly act as a transcriptional repressor by inhibiting the interaction of NR6A1 with the corepressor NCOR1.

Background References

1. Li Y. et. al. USP13 regulates the RAP80-BRCA1 complex dependent DNA damage response. Nat Commun. 2017 Jun 1;8:15752.

2. Lombardi PM. et. al. RAP80, ubiquitin and SUMO in the DNA damage response. J Mol Med (Berl). 2017 Aug;95(8):799-807.

Sequence Similarity

Belongs to the RAP80 family.

Tissue Specificity

Expressed in testis, ovary, thymus and heart. Expressed in germ cells of the testis.

Post-translational Modification

Sumoylated.; Phosphorylated upon DNA damage by ATM or ATR.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q96RL1 Human

SwissProt: Q5U5Q9 Mouse

SwissProt: Q5PQK4 Rat

Synonyms

BRCA1-A complex subunit RAP80 antibody

Nuclear zinc finger protein RAP80 antibody

OTTHUMP00000161441 antibody

OTTHUMP00000223372 antibody

OTTHUMP00000223374 antibody

RAP80 antibody

Receptor associated protein 80 antibody

Receptor-associated protein 80 antibody

Retinoid X receptor interacting protein 110 antibody

Retinoid x receptor interacting protein antibody

Expand

Images

  • Western blot analysis of RAP80 on Siha cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/EM1902-05" style="font-weight: bold;text-decoration: underline;">EM1902-05</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of RAP80 on Siha cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1902-05, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-RAP80 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1902-05" style="font-weight: bold;text-decoration: underline;">EM1902-05</a>, 1/200) for 30 minutes 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 using anti-RAP80 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-05, 1/200) for 30 minutes 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 using anti-RAP80 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1902-05" style="font-weight: bold;text-decoration: underline;">EM1902-05</a>, 1/50) for 30 minutes 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 using anti-RAP80 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-05, 1/50) for 30 minutes 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 RAP80 was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/EM1902-05" style="font-weight: bold;text-decoration: underline;">EM1902-05</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of RAP80 was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-05, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation