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Phospho-RPA2-Ser33 Rabbit mAb (AP1479)

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:Mouse, Rat

Western blot analysis of lysates from NIH/3T3 cells using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at 1:1000 dilution. NIH/3T3 cells were treated by Camptothecin (1 μM) at 37℃ for 1 hour.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

ABclonal:Immunohistochemistry - Phospho-RPA2-Ser33 Rabbit mAb (AP1479)

Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at a dilution of 1:800 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at a dilution of 1:800 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at a dilution of 1:800 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at a dilution of 1:800 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479) at a dilution of 1:800 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - Phospho-RPA2-Ser33 Rabbit mAb (AP1479)

Confocal imaging of paraffin-embedded Mouse testis and Mouse testis treated with λpp tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479, dilution 1:200) 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.

Confocal imaging of paraffin-embedded Mouse kidney and Mouse kidney treated with λpp tissue using Phospho-RPA2-Ser33 Rabbit mAb (AP1479, dilution 1:200) 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	Phospho-RPA2-Ser33 Rabbit mAb
Catalog No.	AP1479
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC66738

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	A synthetic phosphorylated peptide around Ser33 of human RPA2 (NP_002937.1).
Sequence	PAPSQA
Gene ID
Swiss Prot
Synonyms	REPA2; RPA32; RP-A p32; RP-A p34
Calculated MW	29kDa
Observed MW	32kDa

Applications

Reactivity	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 1:500 - 1:1000 IHC-P 1:500 - 1:1000 IF/ICC 1:50-1:200 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 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	NIH/3T3, NIH/3T3+Camptothecin
Cellular location	Nucleus, PML body.