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Product Info Summary
| SKU: | A12720-1 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-POLR3F Antibody Picoband®
SKU/Catalog Number
A12720-1
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-POLR3F Antibody Picoband® catalog # A12720-1. Tested in WB, ICC/IF, FCM, ELISA applications. This antibody reacts with Human, Mouse, Rat. 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
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-POLR3F Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A12720-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human POLR3F recombinant protein (Position: K7-E312). Human POLR3F shares 98.7% amino acid (aa) sequence identity with mouse POLR3F.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
A12720-1 is reactive to POLR3F in Human, Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Observed Molecular Weight
36 kDa
Calculated molecular weight
35.684kDa
Background of POLR3F
DNA-directed RNA polymerase III subunit RPC6 is an enzyme that in humans is encoded by the POLR3F gene. The protein encoded by this gene is one of more than a dozen subunits forming eukaryotic RNA polymerase III (RNA Pol III), which transcribes 5S ribosomal RNA and tRNA genes. This protein has been shown to bind both TFIIIB90 and TBP, two subunits of RNA polymerase III transcription initiation factor IIIB (TFIIIB). Unlike most of the other RNA Pol III subunits, the encoded protein is unique to this polymerase. Alternative splicing results in multiple transcript variants.
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
A12720-1 is guaranteed for ELISA, 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, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human PC-3 whole cell, human Hela whole cell, human A431 whole cell, human U251 whole cell, rat brain tissue, rat RH35 whole cell, mouse brain tissue, mouse NIH/3T3 whole cell
ICC/IF: MCF-7 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of POLR3F using anti-POLR3F antibody (A12720-1).
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 PC-3 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat RH35 whole cell lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse NIH/3T3 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-POLR3F antigen affinity purified polyclonal antibody (Catalog # A12720-1) 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 POLR3F at approximately 36 kDa. The expected band size for POLR3F is at 36 kDa.
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Figure 2. IF analysis of POLR3F using anti-POLR3F antibody (A12720-1) and anti-Beta Tubulin antibody (M01857-3).
POLR3F was detected in immunocytochemical section of MCF-7 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-POLR3F Antibody (A12720-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127)(B) and DyLight®550 Conjugated Goat Anti-Mouse IgG (BA1133)(D) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI(C). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 3. Flow Cytometry analysis of A431 cells using anti-POLR3F antibody (A12720-1).
Overlay histogram showing A431 cells stained with A12720-1 (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-POLR3F Antibody (A12720-1, 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.
Protein Target Info & Infographic
Gene/Protein Information For POLR3F (Source: Uniprot.org, NCBI)
Gene Name
POLR3F
Full Name
DNA-directed RNA polymerase III subunit RPC6
Weight
35.684kDa
Superfamily
eukaryotic RPC34/RPC39 RNA polymerase subunit family
Alternative Names
DNA-directed RNA polymerase III 39 kDa polypeptide; DNA-directed RNA polymerase III subunit F; DNA-directed RNA polymerase III subunit RPC6; DNA-directed RNA polymerases III 39 kDa polypeptide; polymerase (RNA) III (DNA directed) polypeptide F (39 kDa); polymerase (RNA) III (DNA directed) polypeptide F, 39 kDa; RNA polymerase III 39 kDa subunit; RNA polymerase III C39 subunit; RNA polymerase III subunit C6; RPC39MGC13517; RPC6 POLR3F C34, RPC39, RPC6 RNA polymerase III subunit F DNA-directed RNA polymerase III subunit RPC6|RNA polymerase III C39 subunit|RNA polymerase III subunit C6|polymerase (RNA) III (DNA directed) polypeptide F, 39 kDa|polymerase (RNA) III subunit F
*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 POLR3F, check out the POLR3F 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 POLR3F: 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-POLR3F Antibody Picoband® (A12720-1)
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