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mouse anti-Rad51 monoclonal antibody (14B4) 8831

$503.00

Antibody summary

  • Mouse monoclonal to Rad51
  • Suitable for: WB,ICC/IF,IHC-P,IP,IHC,PLA
  • Isotype: IgG2b
  • 100 µg
SKU: 8831parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

mouse

isotype

IgG2b

clonality

monoclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

Rad51

available sizes

100 µg

mouse anti-Rad51 monoclonal antibody (14B4) 8831

antibody
Tested applications
WB,IHC,IHC,ICC/IF
Recommended dilutions
Immunoblotting,

Immunoprecipitation: use at 1-5 ug/mL.

Positive control: T24 bladder carcinoma cells and recombinant fusion protein.
Immunogen
GST fusion protein corresponding to full-length Rad51 (aa 1-338) of human Rad51 expressed in E. coli.
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -70°C. Avoid multiple freeze- thaw cycles.
Storage buffer
PBS, pH 7.4
Purity
affinity purified
Clonality
monoclonal
Isotype
IgG2b
Compatible secondaries
goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody 5486
goat anti-mouse IgG, H&L chain specific, biotin conjugated, Conjugate polyclonal antibody 2685
goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody 7854
goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody, crossabsorbed 1706
goat anti-mouse IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1716
goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1721
Isotype control
Mouse monocolonal IgG2b - Isotype Control
target relevance
Protein names
DNA repair protein RAD51 homolog 1 (HsRAD51) (hRAD51) (RAD51 homolog A)
Gene names
RAD51,RAD51 RAD51A RECA
Protein family
RecA family, RAD51 subfamily
Mass
36966Da
Function
FUNCTION: Plays an important role in homologous strand exchange, a key step in DNA repair through homologous recombination (HR) (PubMed:18417535, PubMed:20348101, PubMed:12205100, PubMed:20231364, PubMed:22325354, PubMed:23754376, PubMed:23509288, PubMed:28575658, PubMed:26681308, PubMed:32640219). Binds to single-stranded DNA in an ATP-dependent manner to form nucleoprotein filaments which are essential for the homology search and strand exchange (PubMed:18417535, PubMed:20348101, PubMed:12205100, PubMed:20231364, PubMed:23754376, PubMed:23509288, PubMed:28575658, PubMed:26681308). Catalyzes the recognition of homology and strand exchange between homologous DNA partners to form a joint molecule between a processed DNA break and the repair template (PubMed:18417535, PubMed:20348101, PubMed:12205100, PubMed:20231364, PubMed:23754376, PubMed:23509288, PubMed:28575658, PubMed:26681308). Recruited to resolve stalled replication forks during replication stress (PubMed:27797818, PubMed:31844045). Part of a PALB2-scaffolded HR complex containing BRCA2 and RAD51C and which is thought to play a role in DNA repair by HR (PubMed:24141787, PubMed:12442171). Plays a role in regulating mitochondrial DNA copy number under conditions of oxidative stress in the presence of RAD51C and XRCC3 (PubMed:20413593). Also involved in interstrand cross-link repair (PubMed:26253028). {ECO:0000269|PubMed:12205100, ECO:0000269|PubMed:12442171, ECO:0000269|PubMed:18417535, ECO:0000269|PubMed:20231364, ECO:0000269|PubMed:20348101, ECO:0000269|PubMed:20413593, ECO:0000269|PubMed:22325354, ECO:0000269|PubMed:23509288, ECO:0000269|PubMed:23754376, ECO:0000269|PubMed:24141787, ECO:0000269|PubMed:26253028, ECO:0000269|PubMed:26681308, ECO:0000269|PubMed:27797818, ECO:0000269|PubMed:28575658, ECO:0000269|PubMed:31844045, ECO:0000269|PubMed:32640219}.
Subellular location
SUBCELLULAR LOCATION: Nucleus {ECO:0000269|PubMed:12442171, ECO:0000269|PubMed:15665856, ECO:0000269|PubMed:18417535, ECO:0000269|PubMed:19783859, ECO:0000269|PubMed:23401855, ECO:0000269|PubMed:23509288, ECO:0000269|PubMed:26681308, ECO:0000269|PubMed:26833090, ECO:0000269|PubMed:9192668}. Cytoplasm {ECO:0000269|PubMed:16215984, ECO:0000269|PubMed:26681308}. Cytoplasm, perinuclear region. Mitochondrion matrix {ECO:0000269|PubMed:20413593}. Chromosome {ECO:0000269|PubMed:22325354, ECO:0000269|PubMed:23401855, ECO:0000269|PubMed:26811421, ECO:0000269|PubMed:27797818, ECO:0000269|PubMed:31844045}. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome {ECO:0000269|PubMed:21276791}. Note=Colocalizes with RAD51AP1 and RPA2 to multiple nuclear foci upon induction of DNA damage (PubMed:20154705). DNA damage induces an increase in nuclear levels (PubMed:20154705). Together with FIGNL1, redistributed in discrete nuclear DNA damage-induced foci after ionizing radiation (IR) or camptothecin (CPT) treatment (PubMed:23754376). Accumulated at sites of DNA damage in a SPIDR-dependent manner (PubMed:23509288). Recruited at sites of DNA damage in a MCM9-MCM8-dependent manner (PubMed:23401855). Recruited at sites of DNA damage following interaction with TOPBP1 in S-phase (PubMed:26811421). Colocalizes with ERCC5/XPG to nuclear foci in S phase (PubMed:26833090). Recruited to stalled replication forks during replication stress by the TONSL-MMS22L complex, as well as ATAD5 and WDR48 in an ATR-dependent manner (PubMed:27797818, PubMed:31844045). {ECO:0000269|PubMed:20154705, ECO:0000269|PubMed:23401855, ECO:0000269|PubMed:23509288, ECO:0000269|PubMed:23754376, ECO:0000269|PubMed:26811421, ECO:0000269|PubMed:26833090, ECO:0000269|PubMed:27797818, ECO:0000269|PubMed:31844045}.
Tissues
TISSUE SPECIFICITY: Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast.
Structure
SUBUNIT: Forms linear homooligomers, giving rise to a RAD51 nucleoprotein filament, which is essential for strand-pairing reactions during DNA recombination. Interacts with BRCA1 and either directly or indirectly with p53. Interacts with XRCC3, RAD54L and RAD54B. Interacts with the BCDX2 subcomplex RAD51C:RAD51B. Component of the homologous recombination repair (HR) complex composed of ERCC5/XPG, BRCA2, PALB2, DSS1 and RAD51 (PubMed:26833090). Interacts directly with PALB2 which may serve as a scaffold for a HR complex containing PALB2, BRCA2, RAD51C, RAD51 and XRCC3. Interacts with RAD51AP1 and RAD51AP2. Interacts with CHEK1, and this may require prior phosphorylation of CHEK1. Interacts with the MND1-PSMC3IP heterodimer. Found in a complex, at least composed of BLM, RAD51 and SPIDR; the complex formation is mediated by SPIDR. Interacts with SPIDR; the interaction is direct and recruits RAD51 to DNA damage sites. Interacts with FIGNL1 (via N-terminal one-half region); the interaction is direct. Interacts with RAD51AP1 (via C-terminal region); the interaction is direct. Interacts with NABP2, RPA1, PALB2 and RAD51. Interacts with SWI5/C9orf119, and at lower level with SFR1/MEIR5. Interacts with hyperphosphorylated RPA2; this interaction is necessary for efficient recruitment to chromatin in response to DNA damage. Interacts with SWSAP1; involved in homologous recombination repair. Interacts with PARPBP, BRCA2 and RECQL5; these interactions interfere with the formation of the RAD51-DNA homologous recombination structure. Interacts with POLQ; POLQ acts as an inhibitor of homology-recombination repair (HR) pathway by limiting RAD51 accumulation at resected ends (PubMed:25642963). Interacts with FBH1 (PubMed:23393192). Interacts with POLN (PubMed:19995904). Interacts with RFWD3 (PubMed:28575658). Interacts with the MCM8-MCM9 complex; the interaction recruits RAD51 to DNA damage sites (PubMed:23401855). Component of a multiprotein complex with MEIOB and SPATA22. Interacts with the complex BRME1:HSF2BP:BRCA2 (By similarity). Interacts with HELQ; stimulating HELQ DNA helicase activity and ability to unwing DNA (PubMed:34937945). Interacts with MMS22L; the interaction is direct and promotes recruitment of RAD51 to sites of DNA damage (PubMed:27797818). Interacts with the ATAD5 RFC-like complex (PubMed:31844045). Within the ATAD5 RFC-like complex, interacts with ATAD5 (via N-terminus); the interaction is direct and enhanced under replication stress (PubMed:31844045). Interacts with WDR48; the interaction is enhanced under replication stress (PubMed:31844045). Interacts with DNA helicase ZGRF1; the interaction promotes RAD51 strand exchange activity (PubMed:32640219). Interacts (when phosphorylated) with TOPBP1; interaction takes place following phosphorylation by CK2 and PLK1 and promotes recruitment to DNA damage sites (PubMed:26811421). {ECO:0000250|UniProtKB:Q08297, ECO:0000269|PubMed:10851248, ECO:0000269|PubMed:11842113, ECO:0000269|PubMed:12427746, ECO:0000269|PubMed:12442171, ECO:0000269|PubMed:14580352, ECO:0000269|PubMed:15665856, ECO:0000269|PubMed:16990250, ECO:0000269|PubMed:18449195, ECO:0000269|PubMed:19995904, ECO:0000269|PubMed:20154705, ECO:0000269|PubMed:20231364, ECO:0000269|PubMed:20348101, ECO:0000269|PubMed:20705237, ECO:0000269|PubMed:20871615, ECO:0000269|PubMed:21252223, ECO:0000269|PubMed:21965664, ECO:0000269|PubMed:22153967, ECO:0000269|PubMed:23393192, ECO:0000269|PubMed:23401855, ECO:0000269|PubMed:23509288, ECO:0000269|PubMed:23754376, ECO:0000269|PubMed:24141787, ECO:0000269|PubMed:25642963, ECO:0000269|PubMed:26811421, ECO:0000269|PubMed:26833090, ECO:0000269|PubMed:27797818, ECO:0000269|PubMed:28575658, ECO:0000269|PubMed:31844045, ECO:0000269|PubMed:32640219, ECO:0000269|PubMed:34937945, ECO:0000269|PubMed:9192668, ECO:0000269|PubMed:9321665, ECO:0000269|PubMed:9396801, ECO:0000269|PubMed:9461559}.
Post-translational modification
PTM: 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 (PubMed:28575658). {ECO:0000269|PubMed:23393192, ECO:0000269|PubMed:28575658}.; PTM: Phosphorylation of Thr-309 by CHEK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination (PubMed:15665856). Phosphorylated at Ser-14 by PLK1, triggering phosphorylation at Thr-13 by CK2, thereby promoting interaction with TOPBP1 and recruitment to DNA damage sites during S-phase (PubMed:22325354, PubMed:26811421). Phosphorylation by ABL1 inhibits function (PubMed:9461559). {ECO:0000269|PubMed:15665856, ECO:0000269|PubMed:22325354, ECO:0000269|PubMed:26811421, ECO:0000269|PubMed:9461559}.
Target Relevance information above includes information from UniProt accession : Q06609
The UniProt Consortium

Data

benchmark-antibodies_anti-rad51_antibody_8831_1.jpg
Immunofluorecent staning of RAD51 nuclear foci in U2OS cells using RAD51 14B4 antibody (8831).
Cells were pre-extracted with CSK buffer before fixation with 4% PFA.
RAD51 14B4 was used at 1:1000 dultion. DAPI was used to counterstain the nucleus.
Scale bar, 10 mciro-meter.
benchmark-antibodies_anti-rad51_antibody_8831_2.jpg
Mouse anti-RAD51 14B4 antibody (8831) was used in IP assay (immunoprecipitation) using HeLa cell extract prepared with lysis 180 buffer (40 mM Tris-HCl pH8.0, 180 mM NaCl, 1 mMEDTA, 0.5% NP-40).
Rabbit anti-RAD51 antibody was used for subsequent WB detection of immunoprecipated RAD51.
Whole cell extract was used as input.
benchmark-antibodies_anti-rad51_antibody_8831_3.jpg
RAD51 antibody 14B4 (8831) detects RAD51 in nucleus on BT483 xenograft by immunohistochemical analysis.
Sample: Paraffin-embedded BT483 xenograft.
RAD51 antibody 14B4 (88318) dilution: 1:200.
Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min
benchmark-antibodies_anti-rad51_antibody_8831_4.jpg
Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Rad51 antibody [14B4] (8831) diluted at 1:500. The HRP-conjugated anti-mouset IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
benchmark-antibodies_anti-rad51_antibody_8831_5.jpg
Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Rad51 antibody [14B4] (8831) diluted at 1:500. The HRP-conjugated anti-mouset IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
benchmark-antibodies_anti-rad51_antibody_8831_6.jpg
Whole cell extract (30 µg) was separated by 10% SDS-PAGE, and the membrane was blotted with Rad51 antibody [14B4] (8831) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
benchmark-antibodies_anti-rad51_antibody_8831_7.jpg
Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Rad51 antibody [14B4] (8831) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
benchmark-antibodies_anti-rad51_antibody_8831_8.jpg
Rad51 antibody [14B4] detects Rad51 protein at nuclear foci by immunofluorescent analysis.
Sample: Mock and treated HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: Rad51 stained by Rad51 antibody [14B4] (8831) diluted at 1:500.
Blue: Fluoroshield with DAPI.

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




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Protocols

relevant to this product
Western blot
IHC
ICC

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