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Product Info Summary
| SKU: | A11611-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-POLR3D Antibody Picoband®
SKU/Catalog Number
A11611-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-POLR3D Antibody Picoband® catalog # A11611-2. Tested in WB, ICC/IF, FCM, ELISA applications. This antibody reacts with Human, 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-POLR3D Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A11611-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human POLR3D recombinant protein (Position: K231-L360). Human POLR3D shares 92.3% amino acid (aa) sequence identity with mouse POLR3D.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
A11611-2 is reactive to POLR3D in Human, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Observed Molecular Weight
44 kDa
Calculated molecular weight
44.396kDa
Background of POLR3D
DNA-directed RNA polymerase III subunit RPC4 is an enzyme that in humans is encoded by the POLR3D gene. This gene complements a temperature-sensitive mutant isolated from the BHK-21 Syrian hamster cell line. It leads to a block in progression through the G1 phase of the cell cycle at nonpermissive temperatures.
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
A11611-2 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, 0.1-0.25 μg/ml, Human, 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 Hela whole cell, human 293T whole cell, human HEL whole cell, human K562 whole cell, rat testis tissue
ICC/IF: MCF-7 cell
FCM: 293T cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of POLR3D using anti-POLR3D antibody (A11611-2).
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 Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human HEL whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat testis tissue 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-POLR3D antigen affinity purified polyclonal antibody (Catalog # A11611-2) at 0.25 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 POLR3D at approximately 44 kDa. The expected band size for POLR3D is at 44 kDa.
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Figure 2. IF analysis of POLR3D using anti-POLR3D antibody (A11611-2) and anti-Beta Tubulin antibody (M01857-3).
POLR3D 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-POLR3D Antibody (A11611-2) 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 293T cells using anti-POLR3D antibody (A11611-2).
Overlay histogram showing 293T cells stained with A11611-2 (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-POLR3D Antibody (A11611-2, 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 POLR3D (Source: Uniprot.org, NCBI)
Gene Name
POLR3D
Full Name
DNA-directed RNA polymerase III subunit RPC4
Weight
44.396kDa
Superfamily
eukaryotic RPC4/POLR3D RNA polymerase subunit family
Alternative Names
BN51 (BHK21) temperature sensitivity complementing; BN51; BN51TDNA-directed RNA polymerase III 47 kDa polypeptide; DNA-directed RNA polymerase III subunit D; DNA-directed RNA polymerase III subunit RPC4; polymerase (RNA) III (DNA directed) polypeptide D, 44kDa; Protein BN51; RNA polymerase III 47 kDa subunit; RNA polymerase III subunit C4; RPC4; RPC53 homolog; temperature sensitive complementation, cell cycle specific, tsBN51; TSBN51RPC53 POLR3D BN51T, C53, RPC4, RPC53, TSBN51 RNA polymerase III subunit D DNA-directed RNA polymerase III subunit RPC4|BN51 (BHK21) temperature sensitivity complementing|DNA-directed RNA polymerase III 47 kDa polypeptide|DNA-directed RNA polymerase III subunit D|RNA polymerase III 47 kDa subunit|RNA polymerase III subunit C4|RPC53 homolog|polymerase (RNA) III (DNA directed) polypeptide D, 44kDa|polymerase (RNA) III subunit D|temperature sensitive complementation, cell cycle specific, tsBN51
*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 POLR3D, check out the POLR3D 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 POLR3D: 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-POLR3D Antibody Picoband® (A11611-2)
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