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Product Info Summary
| SKU: | PA1716 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Replication factor C subunit 1 RFC1 Antibody Picoband®
SKU/Catalog Number
PA1716
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Replication factor C subunit 1 RFC1 Antibody catalog # PA1716. Tested in Flow Cytometry, IF, IHC, 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-Replication factor C subunit 1 RFC1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1716)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human RFC1.
*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
PA1716 is reactive to RFC1 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
150 kDa
Calculated molecular weight
128255 MW
Background of RFC1
RFC (Replication Factor C), also called activator-1, is a multimeric primer-recognition protein consisting of 5 distinct subunits of 145, 40, 38, 37, and 36.5 kD. Human RFC was purified from extracts of HeLa cells as a host factor essential for the in vitro replication of simian virus 40 (SV40) DNA. RFC, in the presence of ATP, assembles proliferating-cell nuclear antigen and DNA polymerase-delta or polymerase-epsilon on primed DNA templates. Tight interfacial coordination of the ATP analog ATP-gamma-S by RFC resulted in a spiral arrangement of the ATPase domains of the clamp loader above the PCNA ring.
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
PA1716 is guaranteed for Flow Cytometry, IF, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human 293T whole cell, human Hela whole cell, human HepG2 whole cell, human Raji whole cell, human K562 whole cell, human CACO-2 whole cell
IHC: human mammary cancer tissue
ICC/IF: U2OS cell
FCM: Hela cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of RFC1 using anti-RFC1 antibody (PA1716).
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 293T whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: human CACO-2 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-RFC1 antigen affinity purified polyclonal antibody (Catalog # PA1716) 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 RFC1 at approximately 150 kDa. The expected band size for RFC1 is at 128 kDa.
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Figure 2. IHC analysis of RFC1 using anti-RFC1 antibody (PA1716).
RFC1 was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-RFC1 Antibody (PA1716) 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. IF analysis of RFC1 using anti-RFC1 antibody (PA1716) and anti-Beta Tubulin antibody (M01857-3).
RFC1 was detected in immunocytochemical section of U2OS cell. 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-RFC1 Antibody (PA1716) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and DyLight®594 Conjugated Goat Anti-Mouse IgG (BA1141) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 4. Flow Cytometry analysis of Hela cells using anti-RFC1 antibody (PA1716).
Overlay histogram showing Hela cells stained with PA1716 (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-RFC1 Antibody (PA1716, 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 (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For RFC1 (Source: Uniprot.org, NCBI)
Gene Name
RFC1
Full Name
Replication factor C subunit 1
Weight
128255 MW
Superfamily
activator 1 large subunit family
Alternative Names
Replication factor C subunit 1;Activator 1 140 kDa subunit;A1 140 kDa subunit;Activator 1 large subunit;Activator 1 subunit 1;DNA-binding protein PO-GA;Replication factor C 140 kDa subunit;RF-C 140 kDa subunit;RFC140;Replication factor C large subunit;RFC1;RFC140; RFC1 A1, CANVAS, MHCBFB, PO-GA, RECC1, RFC40, RFC1 replication factor C subunit 1 replication factor C subunit 1|A1 140 kDa subunit|DNA-binding protein PO-GA|MHC binding factor, beta|RF-C 140 kDa subunit|activator 1 140 kDa subunit|activator 1 large subunit|activator 1 subunit 1|replication factor C (activator 1) 1, 145kDa|replication factor C 140 kDa subunit|replication factor C large subunit|replication factor C1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on RFC1, check out the RFC1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for RFC1: 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].
Specific Publications For Anti-Replication factor C subunit 1 RFC1 Antibody Picoband® (PA1716)
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Customer Q&As
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4 Customer Q&As for Anti-Replication factor C subunit 1 RFC1 Antibody Picoband®
Question
We are currently using anti-RFC1 antibody PA1716 for mouse tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2019-12-16
Answer
The anti-RFC1 antibody (PA1716) has not been validated for cross reactivity specifically with canine tissues, though 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 canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-12-16
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for liver using anti-RFC1 antibody PA1716. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-08-23
Answer
We appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-08-23
Question
We want to test anti-RFC1 antibody PA1716 on rat liver for research purposes, then I may be interested in using anti-RFC1 antibody PA1716 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2019-04-12
Answer
The products we sell, including anti-RFC1 antibody PA1716, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2019-04-12
Question
Will PA1716 anti-RFC1 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2018-01-25
Answer
You can see on the product datasheet, PA1716 anti-RFC1 antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2018-01-25
