Anti-Rad52 (human) antibody, rabbit polyclonal
Product code70-015

Size

50 µg

Storage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityAffinity-purified with immunogen.

Immunogen

Full-size recombinant human Rad52 protein, no tag-peptide attached.

IsotypeRabbit IgG

Reactivity

Human, mouse, rat, hamster

Special notes

N/A

Application

1. Western blotting (1/1,000~1/3,000)
2. Immuno-precipitation (1/50-1/1,000))
3. Immunofluorescence staining (1/1,000)
4. ELISA (Assay dependent)

Background

Human Rad52 protein is a functional and structural homolog of yeast Rad52
protein, which plays a major role in genetic recombination and recombination
repair with the RAD51 recombinase by mediating strand annealing between
homologous DNA strands. Rad52 functionally and physically interacts with
Rad51 in recombination processes. Rad52 foci formation is induced by DNA
damage.

Data Link

UniProtKB P43351 (RAD52_HUMAN)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 70-015 Anti-Rad52 (human) antibody, rabbit polyclonal

123452501501007550372520(kD)

Fig,1 Western Blot of Rad52 in mammalian cells
M: Marker proteins
1: Recombinant human Rad52 protein (0.2 ng)
2: HeLa cell extrat (17 ug)
3. MCF7 cell extract (27 ug)
4. NIH3T3 cell extract (20 ug)
5. CHO cell extract (21 ug).The samples were separated by electrophoresis on 10% SDS-PAGE and blotted onto PVDF membrane.
The anti-Rad52 antibody was used at 1/1,000 dilution and as the second antibody, HRP-conjugatedgoat anti-rabbit IgG (ab97051) was used at 1/10,000 dilution.
Molecular mass of human Rad52 protein is 47 kDa

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Fig.2 Weastern blot of Rad52 in Rat cells
1. Human MCF-7 cell lysate 16 μg
2. Rat PC-12 cell 40 μg
Proteins in the lysates were separated on SDS-PAGE (12.5%) and electro-blotted to PVDF membraneovernight. The blot was blocked with 5% skim milk. Anti-Rad52 antibody was used at 1/1,000 dilution
and as the second antibody, HRP conjugated goat anti-rabbit IgG (ab97051) was used at 1/10,000dilution.

Fig.3 Immunoprecipitation of Rad52 from human cell lysate.
1.Add 10 µg of anti-Rad52 antibody and 50 µg of Dynabeads proteinA
2. Incubate for 10 min at room temperature, wash and resuspend in 100 µl of the same buffer.
3. Add 100 µl (300 µg in PBS) lysate of human MCF7 cells
4.Incubate for and collect the immunocomplex with Dynabeeds magnetic apparatus
5. Elute the immunocomplex and analyze the presence of Rad52 in the supernatant (lane 2) and theeluate (lane 1) by western blotting with anti-Rad52 antibody.
6. As the second antibody, HRP-conjugated goat anti-rabbit IgG VeriBlot for IP (ab131366) was used
at 1/10,000 dilution.
Lane 3 is western blot of MCF’ lysate (16 µg)
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Fig.4

HU-induced focus formation of Rad52.

U2OS 40-2-6 cells were untreated or treated with 2 mM hydroxyurea (HU) for 24 h. The cells werepre-extracted with 0.1% TritonX-100 in PBS, fixed with 3.7% formaldehyde in PBS, andimmunostained with anti-Rad52 antibody (green) at 1/50 dilution, followed by DAPI staining (blue).
The second antibody was CF488A Goat anti-Rabbit IgG (Biotium) used at 1/1,000 dilution.TritonX100 treatment extracts chromatin-free Rad52 from nuclei. Scale bars, 20 μm. The image is by courtesyof Prof .M.Fujita of Kyushu University.

Fig.5. Detection of Ra52 protein in human fibroblast cell line, GM0637 by immunofluorescencestaining
Cells were fixed with 4% paraformaldehyde in 1x PBS for 10 min, washed 3 times with PBS for 3min, permeabilized by treatment with solution containing 1% SDS and 0.5% Triton for 10 min, washed
3 times with PBS for 3 min, incubated with anti-Rad52 antibody for 30 min at 37℃, washed 3 times
with PBS for 3 min, incubated with secondary antibody for 30 min at 37℃, washed 3 times with PBS
for 3 min, stained with DAPI for 1 min and mounted. Photographed with confocal microscope.
Anti-Rad52 antibody was used at 1/1,000 dilution. As the secondary antibody, goat anti-rabbit IgG
conjugated with Alexa 488 was used at 1/2,000 dilution. The pictures were by courtesy of Prof. S.
Tashiro and Mr. A. Fukuto at Hiroshima University.
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Fig.6 Detection of Rad52 protein in HeLa cells and MCF7 cells by immunofluorescence staining
Fixed with 4% paraformaldehyde, and permeabilized with 0.25% TritonX-100. DNA was stained withDAPI. Anti-Rad52 antibody was used at 1/1,000 dilution and second antibody, goat anti-rabbit IgG
Alex 488 at 1/1,000 dilution.

Fig 7 ELISA of human Rad52 protein with anti-Rad52 antibody.
ELISA analysis of human Rad52 protein, using the anti-Rad52 antibody. Plate was coated with 100μl of recombinant human Rad52 protein at 1 μg/ml (BioAcademia 10-003) per well and 100 μl of theanti-Rad52 antibody at the indicated dilution was added to each well and incubated. After washing,goat anti-rabbit-IgG conjugated with HRP was added as a secondary antibody. Color was developedwith TMB as substrate.
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Reference: There has been no publication yet using this antibody.
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