Anti-RFC4 Antibody Picoband™

Product Name

Anti-RFC4 Antibody Picoband™

View all RFC4 Antibodies

SKU/Catalog Number

A07702-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-RFC4 Antibody Picoband™ catalog # A07702-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-RFC4 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A07702-1)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human RFC4 recombinant protein (Position: E42-Q323).

*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

A07702-1 is reactive to RFC4 in Human

Applications

A07702-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

37 kDa

Calculated molecular weight

39.682kDa

Background of RFC4

Replication factor C subunit 4 is a protein that in humans is encoded by the RFC4 gene. The elongation of primed DNA templates by DNA polymerase delta and DNA polymerase epsilon requires the accessory proteins proliferating cell nuclear antigen (PCNA) and replication factor C (RFC). RFC, also named activator 1, is a protein complex consisting of five distinct subunits of 140, 40, 38, 37, and 36 kD. This gene encodes the 37 kD subunit. This subunit forms a core complex with the 36 and 40 kDa subunits. The core complex possesses DNA-dependent ATPase activity, which was found to be stimulated by PCNA in an in vitro system. Alternatively spliced transcript variants encoding the same protein have been reported.

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

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/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, 0.25-0.5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of RFC4 using anti-RFC4 antibody (A07702-1).
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 30 ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human HepG2 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-RFC4 antigen affinity purified polyclonal antibody (Catalog # A07702-1) 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 RFC4 at approximately 37 kDa. The expected band size for RFC4 is at 40 kDa.

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Figure 2. IF analysis of RFC4 using anti-RFC4 antibody (A07702-1) and anti-Beta Tubulin antibody (M01857-3).
RFC4 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-RFC4 Antibody (A07702-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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 3. Flow Cytometry analysis of Raji cells using anti-RFC4 antibody (A07702-1).
Overlay histogram showing Raji cells stained with A07702-1 (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-RFC4 Antibody (A07702-1, 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 RFC4 (Source: Uniprot.org, NCBI)

Alternative Names

A1 37 kDa subunit; A1; Activator 1 37 kDa subunit; Activator 1 subunit 4; MGC27291; replication factor C (activator 1) 4, 37kDa; Replication factor C 37 kDa subunit; replication factor C subunit 4; RFC 37 kDa subunit; RF-C 37 kDa subunit; RFC37replication factor C (activator 1) 4 (37kD) RFC4 A1, RFC37 replication factor C subunit 4 replication factor C subunit 4|A1 37 kDa subunit|RF-C 37 kDa subunit|RFC 37 kDa subunit|activator 1 37 kDa subunit|activator 1 subunit 4|replication factor C (activator 1) 4, 37kDa|replication factor C 37 kDa subunit

For more info on RFC4, check out the RFC4 Infographic

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In this infographic, you will see the following information for RFC4: 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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