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Product Info Summary
| SKU: | A00123-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-BRD4 Antibody Picoband®
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, 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-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.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00123-3 is reactive to BRD4 in Human, Mouse, Rat
Observed Molecular Weight
240 kDa
Calculated molecular weight
152.2 kDa
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.
Application & Images
Applications
A00123-3 is guaranteed for 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
Positive Control
WB: human HepG2 whole cell, human A549 whole cell, human MCF-7 whole cell, rat C6 whole cell, mouse NIH/3T3 whole cell
ICC/IF: MCF-7 cell
FCM: A549 cell
Validation Images & Assay Conditions
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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 HepG2 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: rat C6 whole cell lysates,
Lane 5: 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-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 240 kDa. The expected band size for BRD4 is at 152 kDa.
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IF analysis of BRD4 using anti-BRD4 antibody (A00123-3).
BRD4 was detected in an immunocytochemical section of MCF-7 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 1:500 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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Flow Cytometry analysis of A549 cells using anti-BRD4 antibody (A00123-3).
Overlay histogram showing A549 cells stained with A00123-3 (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-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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-BRD4 Antibody Picoband® (A00123-3)
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