Anti-POLR2B Antibody Picoband™

Product Name

Anti-POLR2B Antibody Picoband™

View all RPB2 Antibodies

SKU/Catalog Number

A08699-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-POLR2B Antibody Picoband™ catalog # A08699-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

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-POLR2B Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A08699-1)

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 POLR2B recombinant protein (Position: E49-Q1145).

*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

A08699-1 is reactive to POLR2B in Human, Mouse, Rat

Applications

A08699-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

130 kDa

Calculated molecular weight

Background of RPB2

DNA-directed RNA polymerase II subunit RPB2 is an enzyme that in humans is encoded by the POLR2B gene. This gene encodes the second largest subunit of RNA polymerase II (Pol II), a DNA-dependent RNA polymerase that catalyzes the transcription of DNA into precursors of mRNA, snRNA and microRNA. This subunit and the largest subunit form opposite sides of the center cleft of Pol II. Deletion of the flap loop region of this subunit results in a decrease in the rate of transcriptional elongation. 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.

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Reconstitution

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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of POLR2B using anti-POLR2B antibody (A08699-1).
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 A431 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Ramos 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-POLR2B antigen affinity purified polyclonal antibody (Catalog # A08699-1) 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 POLR2B at approximately 130 kDa. The expected band size for POLR2B is at 134 kDa.

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Figure 2. IF analysis of POLR2B using anti-POLR2B antibody (A08699-1) and anti-Beta Tubulin antibody (M01857-3).
POLR2B was detected in immunocytochemical section of A549 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-POLR2B Antibody (A08699-1) 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 U251 cells using anti-POLR2B antibody (A08699-1).
Overlay histogram showing U251 cells stained with A08699-1 (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-POLR2B Antibody (A08699-1, 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 POLR2B (Source: Uniprot.org, NCBI)

Alternative Names

DNA-directed RNA polymerase II subunit RPB2; EC 2.7.7; EC 2.7.7.6; hRPB140; hsRPB2; POL2RB; polymerase (RNA) II (DNA directed) polypeptide B (140kD); polymerase (RNA) II (DNA directed) polypeptide B, 140kDa; RNA polymerase II subunit 2; RNA polymerase II subunit B2; RNA polymerase II subunit2, RNA polymerase II second largest subunit POLR2B POL2RB, RPB2, hRPB140 RNA polymerase II subunit B DNA-directed RNA polymerase II subunit RPB2|DNA-directed RNA polymerase II 140 kDa polypeptide|DNA-directed RNA polymerase II subunit B|RNA polymerase II second largest subunit|RNA polymerase II subunit 2|RNA polymerase II subunit B2|polymerase (RNA) II (DNA directed) polypeptide B, 140kDa|polymerase (RNA) II subunit B

For more info on POLR2B, check out the POLR2B Infographic

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In this infographic, you will see the following information for POLR2B: 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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