Phospho-RPA32/RPA2 (S4, S8) Recombinant Rabbit Monoclonal Antibody [PSH24-99]
cat.: HA751963
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-99 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 29 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser4, Ser8 of Human RPA32 / RPA2 aa 1-50 / 270. |
| Positive control: | HeLa (Human cervical adenocarcinoma cell) cell lysate, HeLa treated with 100mJ/cm2 UV then recovery for 2 hours cell lysate, NIH/3T3 treated with 25μM Etoposide for 5 hours cell lysate, C6 treated with 25μM Etoposide for 5 hours cell lysate. |
| Subcellular location: | Nucleus, PML body. |
| Recommended Dilutions:
WB IF-Cell IP |
1:5,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P15927 Human | Q62193 Mouse | Q63528 Rat |
| Alternative names: | 60S acidic ribosomal protein P1 AA409079 AI325195 AU020965 ik:tdsubc_2g1 M(2)21C MGC137236 OTTHUMP00000004008 p32 p34 RCJMB04_6d17 replication protein A2, 32kDa REPA2 Replication factor A protein 2 Replication protein A 32 kDa subunit Replication protein A 32kDa subunit Replication protein A 34 kDa subunit Replication protein A Replication Protein A2 (32kDa) Replication protein A2 Replication protein A2, 32kDa RF-A protein 2 Rf-A2 RFA RFA2_HUMAN RP-A p32 RP-A p34 RP21C RPA 2 RPA 32 RPA Rpa2 RPA32 RPA34 RpLP1 RpP2 xx:tdsubc_2g1 zgc:109822 |
Images
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Fig1:
Western blot analysis of Phospho-RPA32/RPA2 (S4, S8) on different lysates with Rabbit anti-Phospho-RPA32/RPA2 (S4, S8) antibody (HA751963) at 1/5,000 dilution. Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 2: HeLa treated with 100mJ/cm2 UV then recovery for 2 hours cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 6 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751963, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 29 kDa Observed band size: 35 kDa |
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Fig2:
Western blot analysis of Phospho-RPA32/RPA2 (S4, S8) on different lysates with Rabbit anti-Phospho-RPA32/RPA2 (S4, S8) antibody (HA751963) at 1/5,000 dilution. Lane 1: HeLa (Human cervical adenocarcinoma cell) treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate Lane 2: HeLa treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 10 µg/Lane. Exposure time: 1 minute 2 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751963, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 29 kDa Observed band size: 35 kDa |
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Fig3:
Western blot analysis of Phospho-RPA32/RPA2 (S4, S8) on different lysates with Rabbit anti-Phospho-RPA32/RPA2 (S4, S8) antibody (HA751963) at 1/5,000 dilution. Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate Lane 2: NIH/3T3 treated with 25μM Etoposide for 5 hours cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751963, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 29 kDa Observed band size: 35 kDa |
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Fig4:
Western blot analysis of Phospho-RPA32/RPA2 (S4, S8) on different lysates with Rabbit anti-Phospho-RPA32/RPA2 (S4, S8) antibody (HA751963) at 1/5,000 dilution. Lane 1: C6 (Rat glioma cell) cell lysate Lane 2: C6 treated with 25μM Etoposide for 5 hours cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751963, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 29 kDa Observed band size: 35 kDa |
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Fig5:
Application: Immunocytochemistry (IF-cell) Species: Rat Sample: C6 (Rat glioma cell) treated with 25μM Etoposide for 5 hours Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. Antibody dilution buffer: 1% BSA in PBST. Primary antibody: HA751963, 1/1,000, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature. Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). |
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Fig6:
Immunoprecipitation (IP) Phospho-RPA32/RPA2 (S4, S8) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate with HA751963 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751963 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate (input) Lane 2: HA751963 IP in HeLa treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate Lane 3: Rabbit IgG instead of HA751963 in HeLa treated with 100 mJ/cm2 UV, then recovered for 2 hours cell lysate Exposure time: 35 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA751963, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 29 kDa Observed band size: 35 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.