Anti-BRD4 Antibody Picoband™

Product Name

Anti-BRD4 Antibody Picoband™

View all BRD4 Antibodies

SKU/Catalog Number

A00123-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-BRD4 Antibody Picoband™ catalog # A00123-3. Tested in Flow Cytometry, IF, ICC, WB 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-BRD4 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00123-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human BRD4, which shares 95% amino acid (aa) sequence identity with mouse BRD4.

*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

A00123-3 is reactive to BRD4 in Human, Rat

Applications

A00123-3 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

200 kDa

Calculated molecular weight

152.219kDa

Background of BRD4

Bromodomain-containing protein 4 is a protein that in humans is encoded by the BRD4 gene. The protein encoded by this gene is homologous to the murine protein MCAP, which associates with chromosomes during mitosis, and to the human RING3 protein, a serine/threonine kinase. Each of these proteins contains two bromodomains, a conserved sequence motif which may be involved in chromatin targeting. This gene has been implicated as the chromosome 19 target of translocation t(15;19)(q13;p13.1), which defines an upper respiratory tract carcinoma in young people. Two alternatively spliced transcript variants have been described.

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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human, Rat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of BRD4 using anti-BRD4 antibody (A00123-3).
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 293T whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: human PC-3 whole cell lysates,
Lane 6: human HL-60 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-BRD4 antigen affinity purified polyclonal antibody (Catalog # A00123-3) 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 BRD4 at approximately 200 kDa. The expected band size for BRD4 is at 200 kDa.

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Figure 2. IF analysis of BRD4 using anti-BRD4 antibody (A00123-3).
BRD4 was detected in an immunocytochemical section of PC-3 cells. 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-BRD4 Antibody (A00123-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of C6 cells using anti-BRD4 antibody (A00123-3).
Overlay histogram showing C6 cells stained with A00123-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BRD4 Antibody (A00123-3, 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.

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Figure 4. Flow Cytometry analysis of U20S cells using anti-BRD4 antibody (A00123-3).
Overlay histogram showing U20S cells stained with A00123-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BRD4 Antibody (A00123-3, 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 BRD4 (Source: Uniprot.org, NCBI)

Alternative Names

BRD4; bromodomain containing 4; bromodomain-containing 4; CAPchromosome-associated protein; HUNK1; HUNK1bromodomain-containing protein 4; HUNKI; MCAP; Protein HUNK1 BRD4 CAP, HUNK1, HUNKI, MCAP bromodomain containing 4 bromodomain-containing protein 4|chromosome-associated protein|mitotic chromosome-associated protein

For more info on BRD4, check out the BRD4 Infographic

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In this infographic, you will see the following information for BRD4: 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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