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RPA32/RPA2 Rabbit mAb (A26498)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using RPA32/RPA2 Rabbit mAb (A26498) at 1:25000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunohistochemistry - RPA32/RPA2 Rabbit mAb (A26498)

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human colon tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human esophagus tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat testis tissue using RPA32/RPA2 Rabbit mAb (A26498) at a dilution of 1:20000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - RPA32/RPA2 Rabbit mAb (A26498)

Confocal imaging of U-2 OS cells using RPA32/RPA2 Rabbit mAb (A26498, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of HeLa cells using RPA32/RPA2 Rabbit mAb (A26498, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of NIH/3T3 cells using RPA32/RPA2 Rabbit mAb (A26498, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of C6 cells using RPA32/RPA2 Rabbit mAb (A26498, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of paraffin-embedded Human colon cancer tissue using RPA32/RPA2 Rabbit mAb (A26498, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.

Overview

Product name	RPA32/RPA2 Rabbit mAb
Catalog No.	A26498
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC69940

Background

This gene encodes a subunit of the heterotrimeric Replication Protein A (RPA) complex, which binds to single-stranded DNA (ssDNA), forming a nucleoprotein complex that plays an important role in DNA metabolism, being involved in DNA replication, repair, recombination, telomere maintenance, and co-ordinating the cellular response to DNA damage through activation of the ataxia telangiectasia and Rad3-related protein (ATR) kinase. The RPA complex protects single-stranded DNA from nucleases, prevents formation of secondary structures that would interfere with repair, and co-ordinates the recruitment and departure of different genome maintenance factors. The heterotrimeric complex has two different modes of ssDNA binding, a low-affinity and high-affinity mode, determined by which oligonucleotide/oligosaccharide-binding (OB) domains of the complex are utilized, and differing in the length of DNA bound. This subunit contains a single OB domain that participates in high-affinity DNA binding and also contains a winged helix domain at its carboxy terminus, which interacts with many genome maintenance protein. Post-translational modifications of the RPA complex also plays a role in co-ordinating different damage response pathways.

Immunogen information

Immunogen	Recombinant Protein corresponding to a sequence within amino acids 1-270 of human RPA32/RPA2 (NP_002937.1).
Sequence	MWNSGFESYGSSSYGGAGGYTQSPGGFGSPAPSQAEKKSRARAQHIVPCTISQLLSATLVDEVFRIGNVEISQVTIVGIIRHAEKAPTNIVYKIDDMTAAPMDVRQWVDTDDTSSENTVVPPETYVKVAGHLRSFQNKKSLVAFKIMPLEDMNEFTTHILEVINAHMVLSKANSQPSAGRAPISNPGMSEAGNFGGNSFMPANGLTVAQNQVLNLIKACPRPEGLNFQDLKNQLKHMSVSSIKQAVDFLSNEGHIYSTVDDDHFKSTDAE
Gene ID
Swiss Prot
Synonyms	REPA2; RPA32; RP-A p32; RP-A p34
Calculated MW	29kDa
Observed MW	32kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:15000 - 1:60000 IHC-P 1:10000 - 1:40000 IF/ICC 1:1000 - 1:4000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	293T, U-2 OS, NIH/3T3
Cellular location	Nucleus, PML body.