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Catalog Number | orb570308 |
---|---|
Category | Antibodies |
Description | CDK1 Antibody (monoclonal, 2G11) |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 2G11 |
Tested applications | FC, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Mouse IgG2b |
Immunogen | E.coli-derived human CDK1 recombinant protein (Position: L66-M297). Human CDK1 shares 97.8% and 98.3% amino acid (aa) sequence identity with mouse and rat CDK1, respectively. |
Concentration | 0 |
Dilution range | Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Flow Cytometry, 1-3μg/1x106 cells |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 34 kDa |
UniProt ID | P06493 |
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 | Cyclin-dependent kinase 1; CDK1; Cell division con Read more... |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. 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 500μg/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of CDK1 using anti-CDK1 antibody (orb570308). 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: human HEK293 whole cell lysates Lane 2: human A549 whole cell lysates Lane 3: human HepG2 whole cell lysates Lane 4: human THP-1 whole cell lysates Lane 5: human PANC-1 whole cell lysates Lane 6: human SW620 whole cell lysates Lane 7: rat RH35 whole cell lysates Lane 8: mouse NIH/3T3 whole cell lysates. 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 mouse anti-CDK1 antigen affinity purified monoclonal antibody (Catalog # orb570308) 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-mouse 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 # orb90502) with Tanon 5200 system. A specific band was detected for CDK1 at approximately 34KD. The expected band size for CDK1 is at 34KD.
IHC analysis of CDK1 using anti-CDK1 antibody (orb570308). CDK1 was detected in paraffin-embedded section of human colon cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CDK1 Antibody (orb570308) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of CDK1 using anti-CDK1 antibody (orb570308). CDK1 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CDK1 Antibody (orb570308) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of CDK1 using anti-CDK1 antibody (orb570308). CDK1 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CDK1 Antibody (orb570308) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of CDK1 using anti-CDK1 antibody (orb570308). CDK1 was detected in paraffin-embedded section of mouse testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CDK1 Antibody (orb570308) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of CDK1 using anti-CDK1 antibody (orb570308). CDK1 was detected in paraffin-embedded section of rat testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CDK1 Antibody (orb570308) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
Flow Cytometry analysis of PC-3 cells using anti-CDK1 antibody (orb570308). Overlay histogram showing PC-3 cells stained with orb570308 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-CDK1 Antibody (orb570308, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of U20S cells using anti-CDK1 antibody (orb570308). Overlay histogram showing U20S cells stained with orb570308 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-CDK1 Antibody (orb570308, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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