WB result of RPA70 Recombinant Rabbit mAb
Primary antibody: RPA70 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HEK-293 whole cell lysate 20 µg
Lane 2: K562 whole cell lysate 20 µg
Lane 3: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 68 kDa
Observed MW: 68 kDa
RPA70 Recombinant Rabbit mAb (S-1653-9)
RPA70 Recombinant Rabbit mAb (S-1653-9)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | RPA70 |
| Synonyms | Replication protein A 70 kDa DNA-binding subunit; RP-A p70; Replication factor A protein 1 (RF-A protein 1); Single-stranded DNA-binding protein; RPA1; REPA1 |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P27694 |
| Clone Number | S-1653-9 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu, Rt |
| Positive Sample | HEK-293, K562, HeLa, C6 |
| Predicted Reactivity | Or |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Rt |
| IHC-P | 1:50 | Hu, Rt |
| ICC | 1:500 | Hu |
| ICFCM | 1:500 | Hu |
Background
RPA70 is part of the heterotrimeric RPA complex, which is the primary single-stranded DNA (ssDNA) binding protein in eukaryotes. It plays a central role in almost all aspects of DNA replication, repair, and recombination. RPA70 is involved in cellular DNA damage response. It can interact with a SUMO-specific protease SENP6 and is modified by SUMO, which is essential for homologous recombination repair of DNA double-strand breaks. RPA70 undergoes phosphorylation, which is important for mediating activation of the ATR checkpoint and maintaining genomic stability. The N-terminal domain of RPA70 has weak DNA binding activity and is involved in protein interactions and cellular DNA damage response. RPA70 and the RPA complex are also considered promising targets for anti-cancer therapeutics. Inhibition of RPA can abrogate ATR activity and decrease the cellular ssDNA protection threshold, making it a potential strategy for single agent or combination treatment in cancer.
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Western Blot
WB result of RPA70 Recombinant Rabbit mAb
Primary antibody: RPA70 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 68 kDa
Observed MW: 68 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) labelling RPA70 antibody at 1/500 dilution (0.1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-RPA70 antibody was used at 1/50 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cervical squamous cell carcinoma. Anti-RPA70 antibody was used at 1/50 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-RPA70 antibody was used at 1/50 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-RPA70 antibody was used at 1/50 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat testis. Anti-RPA70 antibody was used at 1/50 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- RPA70 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
