RPA70 Recombinant Rabbit Monoclonal Antibody [JB75-32]
Catalog# ET7107-51
RPA70 Recombinant Rabbit Monoclonal Antibody [JB75-32]
-
WB
-
IF-Cell
-
IF-Tissue
-
FC
-
Human
-
Rat
-
unconjugated
Overview
Product Name
RPA70 Recombinant Rabbit Monoclonal Antibody [JB75-32]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human RPA70 aa 22-71 / 616.
Species Reactivity
Human, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, FC
Target Molecular Weight
Predicted band size: 68 kDa
Positive Control
HeLa cell lysate, HEK-293 cell lysate, U-2 OS cell lysate, MCF7 cell lysate, K-562 cell lysate, PC-12 cell lysates, Daudi, C6.
Conjugation
unconjugated
Clone Number
JB75-32
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000-1:5,000
-
IF-Cell
-
1:100-1:500
-
FC
-
1:1,000
Target
Function
The single-stranded-DNA-binding proteins (SSBs) are essential for DNA function in prokaryotic and eukaryotic cells, mitochondria, phages and viruses. Replication protein A (RPA), a highly conserved eukaryotic protein, is a heterotrimeric SSB. RPA plays an important role in DNA replication, recombination and repair. The binding of human RPA (hRPA) to DNA involves molecular polarity in which initial hRPA binding occurs on the 5' side of an ssDNA substrate and then extends in the 3' direction to create a stably bound hRPA. RPA is a major damage-recognition protein involved in the early stages of nucleotide excision repair. It can also play a role in telomere maintenance. The RPA 70 kDa subunit binds to ssDNA and mediates interactions with many cellular and viral proteins. The DNA binding domain lies in the middle of RPA 70 kDa subunit and comprises two structurally homologous subdomains oriented in tandem. RPA contains a conserved four cysteine-type zinc-finger motif, which mediates the transition of RPA-ssDNA interaction to a stable RPA-ssDNA complex in a redox-dependent manner.
Background References
1. Aboussekhra A et al. Mammalian DNA nucleotide excision repair reconstituted with purified protein components. Cell 80:859-868 (1995).
2. He Z et al. RPA involvement in the damage-recognition and incision steps of nucleotide excision repair. Nature 374:566-569 (1995).
Sequence Similarity
Belongs to the replication factor A protein 1 family.
Post-translational Modification
DNA damage-induced 'Lys-63'-linked polyubiquitination by PRPF19 mediates ATRIP recruitment to the RPA complex at sites of DNA damage and activation of ATR. Ubiquitinated by RFWD3 at stalled replication forks in response to DNA damage: ubiquitination by RFWD3 does not lead to degradation by the proteasome and promotes removal of the RPA complex from stalled replication forks, promoting homologous recombination.; Sumoylated on lysine residues Lys-449 and Lys-577, with Lys-449 being the major site. Sumoylation promotes recruitment of RAD51 to the DNA damage foci to initiate DNA repair through homologous recombination. Desumoylated by SENP6.
Subcellular Location
Nucleus.
Synonyms
Dmrpa1 antibody
Drosophila Replication Protein A antibody
DRPA antibody
HSSB antibody
Human single stranded DNA binding protein antibody
MST075 antibody
MSTP075 antibody
p70 antibody
REPA1 antibody
Replication factor A antibody
ExpandImages
-
Western blot analysis of RPA70 on different lysates with Rabbit anti-RPA70 antibody (ET7107-51) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: U-2 OS cell lysate
Lane 4: MCF7 cell lysate
Lane 5: K-562 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 68 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-51) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of RPA70 on PC-12 cell lysates with Rabbit anti-RPA70 antibody (ET7107-51) at 1/5,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 68 kDa
Exposure time: 16 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-51) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of Daudi cells labeling RPA70 with Rabbit anti-RPA70 antibody (ET7107-51) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RPA70 antibody (ET7107-51) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling RPA70 with Rabbit anti-RPA70 antibody (ET7107-51) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RPA70 antibody (ET7107-51) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of Daudi cells labeling RPA70.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-51, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of C6 cells labeling RPA70.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-51, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"