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FCM (Flow Cytometry) (Figure 7. Flow Cytometry analysis of JK cells using anti-RPA32/RPA2 antibody (AAA19693).Overlay histogram showing JK cells stained with AAA19693 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-RPA32/RPA2 Antibody (AAA19693, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Mouse anti-Human RPA32/RPA2 Monoclonal Antibody | anti-RPA2 antibody

Anti-RPA32/RPA2 Antibody Picoband (monoclonal, 3B2E9)

Gene Names
RPA2; REPA2; RPA32
Reactivity
Human
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay
Purity
Immunogen affinity purified.
Synonyms
RPA32/RPA2, Antibody; Anti-RPA32/RPA2 Antibody Picoband (monoclonal, 3B2E9); anti-RPA2 antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
Mouse IgG2a
Clone Number
3B2E9
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-RPA2 antibody
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS)
Application Notes
WB: 0.25-0.5 ug/ml, Human
IHC-P: 2-5 ug/ml, Human
ICC/IF: 5 ug/ml, Human
FC/FACS: 1-3 ug/1x10^6 cells, Human
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Mouse IgG for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit for IHC(P) and ICC.
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human RPA32/RPA2 recombinant protein (Position: Q34-H254).
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 7. Flow Cytometry analysis of JK cells using anti-RPA32/RPA2 antibody (AAA19693).Overlay histogram showing JK cells stained with AAA19693 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-RPA32/RPA2 Antibody (AAA19693, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 7. Flow Cytometry analysis of JK cells using anti-RPA32/RPA2 antibody (AAA19693).Overlay histogram showing JK cells stained with AAA19693 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-RPA32/RPA2 Antibody (AAA19693, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 6. IF analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in an immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. DyLight594 Conjugated Goat Anti-Mouse IgG (BA1141) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The tissue section was developed using Phalloidin-iFluor 488 Conjugated. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence) (Figure 6. IF analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in an immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. DyLight594 Conjugated Goat Anti-Mouse IgG (BA1141) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The tissue section was developed using Phalloidin-iFluor 488 Conjugated. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 5. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 4. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 3. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 2. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).RPA32/RPA2 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml mouse anti-RPA32/RPA2 Antibody (AAA19693) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human U20S whole cell lysates,Lane 2: human HEK293 whole cell lysates,Lane 3: human Jurkat whole cell lysates,Lane 4: human K562 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-RPA32/RPA2 antigen affinity purified monoclonal antibody (#AAA19693) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for RPA32/RPA2 at approximately 32 kDa. The expected band size for RPA32/RPA2 is at 32 kDa.)

WB (Western Blot) (Figure 1. Western blot analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (AAA19693).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human U20S whole cell lysates,Lane 2: human HEK293 whole cell lysates,Lane 3: human Jurkat whole cell lysates,Lane 4: human K562 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-RPA32/RPA2 antigen affinity purified monoclonal antibody (#AAA19693) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for RPA32/RPA2 at approximately 32 kDa. The expected band size for RPA32/RPA2 is at 32 kDa.)
Related Product Information for anti-RPA2 antibody
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.
Product Categories/Family for anti-RPA2 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
29,247 Da
NCBI Official Full Name
replication protein A 32 kDa subunit
NCBI Official Synonym Full Names
replication protein A2, 32kDa
NCBI Official Symbol
RPA2
NCBI Official Synonym Symbols
REPA2; RPA32
NCBI Protein Information
replication protein A 32 kDa subunit; RP-A p32; RP-A p34; RF-A protein 2; replication factor A protein 2; replication protein A 34 kDa subunit
UniProt Protein Name
Replication protein A 32 kDa subunit
UniProt Gene Name
RPA2
UniProt Synonym Gene Names
REPA2; RPA32; RPA34; RP-A p32; RF-A protein 2; RP-A p34
UniProt Entry Name
RFA2_HUMAN

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Product Notes

The RPA2 rpa2 (Catalog #AAA19693) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-RPA32/RPA2 Antibody Picoband (monoclonal, 3B2E9) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's RPA32/RPA2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS). WB: 0.25-0.5 ug/ml, Human IHC-P: 2-5 ug/ml, Human ICC/IF: 5 ug/ml, Human FC/FACS: 1-3 ug/1x10^6 cells, Human Tested Species: In-house tested species with positive results. Enhanced Chemiluminescent Kit with anti-Mouse IgG for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit for IHC(P) and ICC. Researchers should empirically determine the suitability of the RPA2 rpa2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "RPA32/RPA2, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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