MA5-14419

antibody from Invitrogen Antibodies

Targeting: RAD51 BRCC5, FANCR, HsRad51, HsT16930, RAD51A, RECA

Provider product page for MA5-14419

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [28]
Comments [0]
Validations
Immunocytochemistry [3]
Other assay [6]

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Product number: MA5-14419 - Provider product page
Provider: Invitrogen Antibodies
Product name: RAD51 Monoclonal Antibody (51RAD01 (3C10))
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14419 targets RAD51 in IHC (P), IP, ICC/IF and WB applications and shows reactivity with Human, mouse, and Rat samples. The MA5-14419 immunogen is recombinant Rad51 protein.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 51RAD01 (3C10)
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

RAD51 protein structure - MA5-14419 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of RAD51 (green) showing staining in the cytoplasm and nucleus of H1299 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a RAD51 monoclonal antibody (Product # MA5-14419) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of RAD51 (green) showing staining in the cytoplasm and nucleus of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a RAD51 monoclonal antibody (Product # MA5-14419) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of RAD51 (green) showing staining in the cytoplasm and nucleus of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a RAD51 monoclonal antibody (Product # MA5-14419) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of RAD51 using RAD51 Monoclonal Antibody (Product # MA5-14419) on Native Human LS174T Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 F133V SPOP impairs cell ability to recover from radiation-induced DNA damage. ( A ) (Left panel) Representative immunofluorescence microphotographs of nuclear gammaH2AX foci in DU145 WT or F133V SPOP cells at 0, 1 and 8 h upon 6 Gy irradiation. (Right panel) Quantification of gammaH2AX foci in WT or F133V SPOP DU145 cells, expressed as mean percentage of cells containing >10 gammaH2AX foci at 0, 1 and 8 h upon irradiation. Data are reported as mean +- SD values. The level of significance was represented as *** p < 0.001, Student's t -test. ( B ) (Left panel) Representative immunofluorescence microphotographs of RAD51 nuclear foci in DU145 WT or F133V SPOP cells at 0 and 8 h upon irradiation (6 Gy). (Right panel) Quantification of RAD51 foci expressed as mean percentage of cells containing >10 RAD51 foci at 0 and 8 h upon irradiation (IR). Data are reported as mean +- SD. ( C ) Western blot analysis and relative quantification of RAD51 protein levels in nucleus/cytosol fractions of DU145 WT or F133V cells at 0 and 8 h upon irradiation (6 Gy). HDAC2 and beta-tubulin were used as equal protein loading controls for nuclear and cytosolic fractions, respectively. Relative protein expression was calculated as nuclear/cytosolic ratio of normalized RAD51 levels.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 SPOP knockdown impairs HR via RAD51 and CHK1 downregulation. ( A ) qRT-PCR detection of RAD51 and CHEK1 transcript levels in DU145 (upper panel) or PC-3 (lower panel) cells at 48 h upon siSPOP transfection, compared to controls, normalized to GAPDH. Data are reported as relative quantity (RQ) +- SD with respect to siNeg transfectants. ( B ) Western blot analysis and relative quantification of RAD51 and CHK1 protein levels in DU145 and PC-3 cells at 48 h upon siSPOP transfection. Vinculin was used as equal protein loading control. ( C ) Representative immunofluorescence microphotographs of nuclear RAD51 foci (cell nuclei: blue; RAD51 foci: green) in DU145 (upper panel) or PC-3 (lower panel) cells at 48 h upon transfection with siSPOP at 0, 0.5 and 6 h after exposure to 6 Gy irradiation and relative quantification, expressed as mean percentage of cells containing >10 RAD51 foci at 0, 0.5 and 6 h after exposure to 6 Gy irradiation. Data are reported as mean +- SD values from three independent experiments. The level of significance was represented as ** p < 0.01, *** p < 0.001, Student's t -test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Expression of Dmc1p and Rad51p. (A) Expression pattern of DMC1 as visualized by RT-PCR. While there is no transcription in vegetatively (veg) growing and starving (S) cells, DMC1 mRNA is maximally abundant in early meiotic cells from 2-4 h after meiosis induction. Timing of expression is the same as that of SPO11 . (B) Southern hybridization with a probe adjacent to the 3'end of the DMC1 locus. Digestion of genomic DNA from knockout, wild-type and DMC1-mCherry strains with EcoRI, which differentially cuts internally in the knockout and tagging constructs, produced the detected fragments of 2.1 kb, 2.8 kb, and 1.8 kb, respectively. The absence of the 2.8 kb fragment in knockout samples indicates that the replacement of the expressed wild-type sequences in the somatic nucleus by the knockout cassette is complete in several lines of both mating types. Also shown is the complete replacement of wild-type DMC1 by a version fused to a sequence encoding the mCherry fluorescent marker. (C) Western detection of Rad51p and Dmc1p with an antibody that recognizes both proteins. At t = 3.5 h after meiosis induction, both Rad51p and Dmc1p are expressed in wild type conjugating cells. In the dmc1 Delta strain, no detectable amount of Dmc1p was expressed 3.5 h after meiosis induction. In a crossing of two rad51 i strains, expression of Rad51p was strongly reduced. Positions of 35 kDa and 40 kDa marker bands are indicated.