|Product Name||Anti-RFC1 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Replication factor C subunit 1(RFC1) detection. Tested with WB, IHC-P in Human.|
|Cite This Product||Anti-RFC1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1716)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human RFC1(46-64aa NSSRKEDDFKQKQPSKKKR).|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Western blot, 0.1-0.5μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
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 50ug of sample under reducing conditions.
Lane 1: HELA Cell Lysate,
Lane 2: SMMC Cell Lysate,
Lane 3: JURKAT Cell Lysate,
Lane 4: CCRF-CEM Cell Lysate,
Lane 5: 293T Cell Lysate.
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- RFC1 antigen affinity purified polyclonal antibody (Catalog # PA1716) at 0.5 Î¼g/mL overnight at 4Â°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 1:10000 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 150KD. The expected band size for RFC1 is at 128KD.
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 1Î¼g/ml rabbit anti- RFC1 Antibody (PA1716) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Replication factor C subunit 1|
|Tissue Specificity||Wide tissue distribution. Undetectable in placental tissue.|
|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;|
|Molecular Weight||128255 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||The elongation of primed DNA templates by DNA polymerase delta and epsilon requires the action of the accessory proteins PCNA and activator 1. This subunit binds to the primer-template junction. Binds the PO-B transcription element as well as other GA rich DNA sequences. Could play a role in DNA transcription regulation as well as DNA replication and/or repair. Can bind single- or double-stranded DNA. .|
*You can search these to find other products in these research areas.
|Background||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.|
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at firstname.lastname@example.org for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact email@example.com
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?A: Some common names include but are not limited to a1 antibody, rfc1 antibody