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Catalog Number | orb654312 |
---|---|
Category | Antibodies |
Description | BRD2 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, WB |
Reactivity | Human |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human BRD2 recombinant protein (Position: M1-Q214). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.25-0.5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 110 kDa |
UniProt ID | P25440 |
Storage | Store 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 freeze-thaw cycles. |
Alternative names | Bromodomain-containing protein 2; O27.1.1; Really Read more... |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of BRD2 using anti-BRD2 antibody (orb654312). 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 A431 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human 293T 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-BRD2 antigen affinity purified polyclonal antibody (Catalog # orb654312) 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 # orb90503) with Tanon 5200 system. A specific band was detected for BRD2 at approximately 110 kDa. The expected band size for BRD2 is at 88 kDa.
IF analysis of BRD2 and Tubulin beta using anti-BRD2 antibody (orb654312) and anti-Tubulin beta antibody (orb610920). BRD2 and Tubulin beta was detected in an immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-BRD2 Antibody (orb654312) and mouse anti-Tubulin beta Antibody (orb610920) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and DyLight?594 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of 293T cells using anti-BRD2 antibody (orb654312). Overlay histogram showing 293T cells stained with orb654312 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BRD2 Antibody (orb654312, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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