Anti-RPA32/RPA2 Antibody Picoband™

Product Name

Anti-RPA32/RPA2 Antibody Picoband™

View all RPA2 Antibodies

SKU/Catalog Number

A02067-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-RPA32/RPA2 Antibody Picoband™ catalog # A02067-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-RPA32/RPA2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02067-1)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human RPA32/RPA2 recombinant protein (Position: Q34-H254).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A02067-1 is reactive to RPA2 in Human

Applications

A02067-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

29 kDa

Calculated molecular weight

29.247kDa

Background of RPA2

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 4μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (A02067-1).
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 50ug of sample under reducing conditions.
Lane 1: human U20S whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human Raji whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-RPA32/RPA2 antigen affinity purified polyclonal antibody (Catalog # A02067-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for RPA32/RPA2 at approximately 29KD. The expected band size for RPA32/RPA2 is at 29KD.

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Figure 2. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (A02067-1).
RPA32/RPA2 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-RPA32/RPA2 Antibody (A02067-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (A02067-1).
RPA32/RPA2 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-RPA32/RPA2 Antibody (A02067-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. IF analysis of RPA32/RPA2 using anti-RPA32/RPA2 antibody (A02067-1).
RPA32/RPA2 was detected in 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 4μg/mL rabbit anti-RPA32/RPA2 Antibody (A02067-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 5. Flow Cytometry analysis of U937 cells using anti-RPA32/RPA2 antibody (A02067-1).
Overlay histogram showing U937 cells stained with A02067-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RPA32/RPA2 Antibody (A02067-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For RPA2 (Source: Uniprot.org, NCBI)

Alternative Names

REPA2; Replication factor A protein 2; replication protein A 32 kDa subunit; Replication protein A 34 kDa subunit; replication protein A2 (32kD); replication protein A2, 32kDa; RF-A protein 2; RP-A p32; RP-A p34; RPA2; RPA32; RPA34 RPA2 REPA2, RP-A p32, RP-A p34, RPA32 replication protein A2 replication protein A 32 kDa subunit|RF-A protein 2|replication factor A protein 2|replication protein A 34 kDa subunit

For more info on RPA2, check out the RPA2 Infographic

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In this infographic, you will see the following information for RPA2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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