Anti-RPB5/POLR2E Antibody Picoband™

Product Name

Anti-RPB5/POLR2E Antibody Picoband™

View all POLR2E Antibodies

SKU/Catalog Number

A08765-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-RPB5/POLR2E Antibody Picoband™ catalog # A08765-1. Tested in ELISA, WB, IF, ICC, Flow Cytometry applications. This antibody reacts with Human, 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-RPB5/POLR2E Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A08765-1)

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 RPB5/POLR2E recombinant protein (Position: Q95-Q210).

*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

A08765-1 is reactive to POLR2E in Human, Rat

Applications

A08765-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

23 kDa

Calculated molecular weight

24.551kDa

Background of POLR2E

DNA-directed RNA polymerases I, II, and III subunit RPABC1 is a protein that in humans is encoded by the POLR2E gene. This gene encodes the fifth largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. This subunit is shared by the other two DNA-directed RNA polymerases and is present in two-fold molar excess over the other polymerase subunits. An interaction between this subunit and a hepatitis virus transactivating protein has been demonstrated, suggesting that interaction between transcriptional activators and the polymerase can occur through this subunit. A pseudogene is located on chromosome 11. Three transcript variants encoding two different isoforms have been found for this gene.

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, Rat
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 RPB5/POLR2E using anti-RPB5/POLR2E antibody (A08765-1).
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 HepG2 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat liver 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-RPB5/POLR2E antigen affinity purified polyclonal antibody (Catalog # A08765-1) 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 RPB5/POLR2E at approximately 23 kDa. The expected band size for RPB5/POLR2E is at 23-25 kDa.

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Figure 2. IF analysis of RPB5/POLR2E using anti-RPB5/POLR2E antibody (A08765-1) and anti-Beta Tubulin antibody (M01857-3).
RPB5/POLR2E 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-RPB5/POLR2E Antibody (A08765-1) 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 Hela cells using anti-RPB5/POLR2E antibody (A08765-1).
Overlay histogram showing Hela cells stained with A08765-1 (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-RPB5/POLR2E Antibody (A08765-1, 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 POLR2E (Source: Uniprot.org, NCBI)

Alternative Names

DNA directed RNA polymerase II 23 kda polypeptide; DNA-directed RNA polymerase II 23 kDa polypeptide; DNA-directed RNA polymerase II subunit E; DNA-directed RNA polymerases I, II, and III subunit RPABC1; hRPB25; hsRPB5; polymerase (RNA) II (DNA directed) polypeptide E (25kD); polymerase (RNA) II (DNA directed) polypeptide E, 25kDa; RPABC1; RPB5 homolog; RPB5; XAP4RNA polymerases I, II, and III subunit ABC1 POLR2E RPABC1, RPB5, XAP4, hRPB25, hsRPB5 RNA polymerase II, I and III subunit E DNA-directed RNA polymerases I, II, and III subunit RPABC1|DNA directed RNA polymerase II 23 kda polypeptide|DNA-directed RNA polymerase II 23 kDa polypeptide|DNA-directed RNA polymerase II subunit E|DNA-directed RNA polymerase subunit RPABC1|RNA polymerase II subunit E|RNA polymerases I, II, and III subunit ABC1|RPB5 homolog|polymerase (RNA) II (DNA directed) polypeptide E, 25kDa|polymerase (RNA) II subunit E

For more info on POLR2E, check out the POLR2E Infographic

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In this infographic, you will see the following information for POLR2E: 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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