Anti-Replication factor C subunit 1 RFC1 Antibody

Product Name

Anti-Replication factor C subunit 1 RFC1 Antibody

View all RFC1 Antibodies

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.

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 (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

Applications

PA1716 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

150 kDa

Calculated molecular weight

128.255kDa

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.

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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, 0.1-0.5μg/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

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.

Gene/Protein Information For RFC1 (Source: Uniprot.org, NCBI)

Alternative Names

A1 140 kDa subunit; A1; Activator 1 140 kDa subunit; Activator 1 large subunit; Activator 1 subunit 1; MGC51786; MHC binding factor, beta; MHCBFB; PO-GA; RECC1; replication factor C (activator 1) 1 (145kD); replication factor C (activator 1) 1, 145kDa; Replication factor C 140 kDa subunit; Replication factor C large subunit; replication factor C subunit 1; replication factor C1; RF-C 140 kDa subunit; RFC; RFC1; RFC140; RFC140DNA-binding protein PO-GA 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

For more info on RFC1, check out the RFC1 Infographic

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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].

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