|Application:||Flow Cytometry, IHC, ICC, WB|
Data & Images
|Product Name||Anti-Cyclin B1 Picoband™ Antibody|
|Description||Rabbit IgG polyclonal antibody for G2/mitotic-specific cyclin-B1(CCNB1) detection. Tested with WB, IHC-F, ICC, FCM in Human.|
|Cite This Product||Anti-Cyclin B1 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9104)|
|Replacement Item||This antibody may replace the following items: sc-130591|sc-166757|sc-245|sc-245-X|sc-293098|sc-53236|sc-53238|sc-594|sc-595|sc-70898|sc-7393|sc-752 from Santa Cruz Biotechnology.|
|Application||Flow Cytometry, IHC, ICC, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
|Immunogen||E.coli-derived human Cyclin B1 recombinant protein (Position: M1-V433). Human Cyclin B1 shares 86% and 85% amino acid (aa) sequences identity with mouse and rat Cyclin B1, respectively.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
*carrier free antibody available upon request.
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||G2/mitotic-specific cyclin-B1|
|Molecular Weight||48337 MW|
|Protein Function||Essential for the control of the cell cycle at the G2/M (mitosis) transition. .|
|Sequence Similarities||Belongs to the cyclin family. Cyclin AB subfamily.|
|Subcellular Localization||Cytoplasm. Nucleus. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome.|
|Alternative Names||G2/mitotic-specific cyclin-B1;CCNB1;CCNB;|
|Research Areas|||cell biology|proteolysis / ubiquitin|proteasome / ubiquitin|ubiquitin e3 enzymes|ring finger e3 ligase| cell biology|cell cycle|cell division|other cell division antibodies||
Background for G2/mitotic-specific cyclin-B1
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Cyclin B1 Picoband™ Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Western blot, 0.1-0.5μg/ml, Human|
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Cyclin B1 Picoband™ Antibody Images
Click the images to enlarge.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
lane 1: recombinant human cyclin b1 protein 0.5ng.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Cyclin B1 antigen affinity purified polyclonal antibody (Catalog # PB9104) 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 1:10000 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 Cyclin B1 at approximately 39KD. The expected band size for Cyclin B1 is at 39KD.
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 50ug of sample under reducing conditions.
lane 1: HELA whole cell lysate ,
lane 2: 293T whole cell lysate ,
lane 3: MCF-7 whole cell lysate ,
lane 4: COLO320 whole cell lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Cyclin B2 antigen affinity purified polyclonal antibody (Catalog # PB9104) 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 1:10000 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 Cyclin B2 at approximately 56KD. The expected band size for Cyclin B2 is at 48KD.
Overlay histogram showing THP-1 cells stained with PB9104 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CCNB1 Antibody (PB9104,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,