Anti-RPA32/RPA2 antibody [9H8] ab2175 is a mouse monoclonal antibody that is used in RPA32/RPA2 western blotting, IHC and flow cytometry. Suitable for human samples.
- Antibody clone 9H8 is the most widely used clone for RPA32/RPA2 on the market and is cited in >370 publications
IgG1
Mouse
pH: 7.3 - 7.5
Preservative: 0.05% Sodium azide
Constituents: 1% BSA
Liquid
Monoclonal
IHC-P | Flow Cyt | WB | |
---|---|---|---|
Human | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50.00000 - 1/100.00000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg for 106 Cells | Notes ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
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As part of the heterotrimeric replication protein A complex (RPA/RP-A), binds and stabilizes single-stranded DNA intermediates that form during DNA replication or upon DNA stress. It prevents their reannealing and in parallel, recruits and activates different proteins and complexes involved in DNA metabolism. Thereby, it plays an essential role both in DNA replication and the cellular response to DNA damage. In the cellular response to DNA damage, the RPA complex controls DNA repair and DNA damage checkpoint activation. Through recruitment of ATRIP activates the ATR kinase a master regulator of the DNA damage response. It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage. Also recruits to sites of DNA damage proteins like XPA and XPG that are involved in nucleotide excision repair and is required for this mechanism of DNA repair. Also plays a role in base excision repair (BER) probably through interaction with UNG. Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. May also play a role in telomere maintenance. RPA stimulates 5'-3' helicase activity of BRIP1/FANCJ (PubMed:17596542).
REPA2, RPA32, RPA34, RPA2, Replication protein A 32 kDa subunit, RP-A p32, Replication factor A protein 2, Replication protein A 34 kDa subunit, RF-A protein 2, RP-A p34
Anti-RPA32/RPA2 antibody [9H8] ab2175 is a mouse monoclonal antibody that is used in RPA32/RPA2 western blotting, IHC and flow cytometry. Suitable for human samples.
- Antibody clone 9H8 is the most widely used clone for RPA32/RPA2 on the market and is cited in >370 publications
IgG1
Mouse
pH: 7.3 - 7.5
Preservative: 0.05% Sodium azide
Constituents: 1% BSA
Liquid
Monoclonal
9H8
IgG fraction
kappa
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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Full details and terms and conditions can be found here:
Terms & Conditions.
Anti-RPA/32/RPA2 antibody [9H8] (ab2175) is used to measure DNA Double-Stranded Breaks resection in U2OS cells +/- Camptotechin (CPT).
ab2175 is used at 1/200 dilution in 1xPBS + 0.2% Triton X-100 for 1 hour at 25°C.
Secondary antibody: anti-mouse Molecular Probes Alexa Fluor® 488 Conjugated at 1/200 dilution.
Formalin-fixed, paraffin-embeded human tonsil tissue stained for RPA32/RPA2 using ab2175 at 1/50 dilution in immunohistochemical analysis. Antigen retrieval with citrate buffer pH 6.0
Overlay histogram showing HeLa cells stained with ab2175 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2175, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.
All lanes: Western blot - Anti-RPA32/RPA2 antibody [9H8] (ab2175) at 5 µg/mL
Lane 1: TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate at 10 µg
Lane 2: SK N BE (Human neuroblastoma) Whole Cell Lysate at 10 µg
Lane 3: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 29 kDa
Observed band size: 32 kDa
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