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Product Info Summary
| SKU: | PB9886 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-RPA70/RPA1 Antibody Picoband®
SKU/Catalog Number
PB9886
PB0931 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-RPA70/RPA1 Antibody Picoband® catalog # PB9886. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-RPA70/RPA1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9886)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human RPA70, different from the related mouse sequence by three amino acids.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
PB9886 is reactive to RPA1 in Human
Observed Molecular Weight
70 kDa
Calculated molecular weight
68.1 kDa
Background of RPA1
Replication protein A 70 kDa DNA-binding subunit is a protein that in humans is encoded by the RPA1 gene. This gene is mapped to chromosome 17p13.3. Replication protein A (RPA) is a heterotrimeric single-strand DNA (ssDNA)-binding protein essential for DNA replication, repair, and recombination. It is composed of 70-kD (RPA1), 32-kD (RPA2), and 14-kD (RPA3) subunits. The RPA1 subunit is responsible for high-affinity ssDNA binding. The RPA complex was originally isolated as a factor essential for in vitro replication of the papovavirus SV40. It had been found that recombinant human RPA1, purified from bacteria, exhibited ssDNA-binding activity comparable to that of the complete RPA complex. RPA1 could substitute for the complete complex in stimulating the activity of DNA polymerase alpha-primase, but it could not substitute for the complete complex in SV40 DNA replication in vitro, suggesting an important functional role for the other subunits.
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.
Application & Images
Applications
PB9886 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
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, 0.1-0.5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human K562 whole cell, human HepG2 whole cell, human 293T whole cell
ICC/IF: A549 cell
FCM: U251 cell
Validation Images & Assay Conditions
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Western blot analysis of RPA70 using anti-RPA70 antibody (PB9886).
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 Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human 293T 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 rabbit anti-RPA70 antigen affinity purified polyclonal antibody (Catalog # PB9886) 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 RPA70 at approximately 70 kDa. The expected band size for RPA70 is at 68 kDa.
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IF analysis of RPA70 using anti-RPA70 antibody (PB9886).
RPA70 was detected in an immunocytochemical section of A549 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 μg/mL rabbit anti-RPA70 Antibody (PB9886) 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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Flow Cytometry analysis of U251 cells using anti-RPA70 antibody (PB9886).
Overlay histogram showing U251 cells stained with PB9886 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RPA70 Antibody (PB9886, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-RPA70/RPA1 Antibody Picoband® (PB9886)
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1 Customer Q&As for Anti-RPA70/RPA1 Antibody Picoband®
Question
We are currently using anti-RPA70/RPA1 antibody PB9886 for human tissue, and we are content with the WB results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on bovine tissues as well?
Verified Customer
Verified customer
Asked: 2019-09-05
Answer
The anti-RPA70/RPA1 antibody (PB9886) has not been tested for cross reactivity specifically with bovine tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-09-05
