Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22956-37] to Cdk9 - ChIP Grade – BSA and Azide free
  • Suitable for: IP, WB, IHC-P, ICC/IF, Flow Cyt, ChIP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free
    See all Cdk9 primary antibodies

  • Description

    Rabbit monoclonal [EPR22956-37] to Cdk9 - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Mouse
    Flow Cyt
    Human
    ICC/IF
    Mouse
    IHC-P
    Mouse
    IP
    Mouse
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T, HeLa, RAW 264.7, PC-12, NIH/3T3 and C6 whole cell lysate; Mouse brain and rat lung tissue lysate; IHC-P: Human pancreas, pancreatic cancer, mouse and rat liver tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow: HeLa and NIH/3T3 cells. IP: NIH/3T3 cell lysate. ChIP: Chromatin from MEF and HeLa cells.

  • General notes

    Ab259268 is the carrier-free version of ab239364. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab259268 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Immunoprecipitation - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunoprecipitation - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    CDK9 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug with ab239364 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239364 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug

    Lane 2: ab239364 IP in NIH/3T3 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239364 in NIH/3T3 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 6 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling CDK9 with ab239364 at 1/2000 dilution (0.30 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat liver (PMID: 9766517, 11282025). The section was incubated with ab239364 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • ChIP - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    ChIP - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    This data was developed using the same antibody clone in a different buffer formulation (ab239364).

    Chromatin was prepared from MEF cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 5 µg of ab239364 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G Sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

    Primers and probes are from paper PMC4103662 (PMID: 23663783)
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CDK9 with ab239364 at 1/2000 dilution (0.30 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse liver (PMID: 9766517, 11282025) The section was incubated with ab239364 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunohistochemical analysis of paraffin-embedded Human pancreatic cancer tissue labeling CDK9 with ab239364 at 1/2000 dilution (0.30 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human pancreatic cancer (PMID: 28231737) The section was incubated with ab239364 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling CDK9 with ab239364 at 1/2000 dilution (0.30 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human pancreas (PMID: 28231737) The section was incubated with ab239364 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunocytochemistry/ Immunofluorescence - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CDK9 with ab239364 at 1/100 (6 ug/ml) dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab239364 anti-CDK9 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Immunocytochemistry/ Immunofluorescence - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CDK9 with ab239364 at 1/100 (6 ug/ml) dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab239364 anti-CDK9 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Flow Cytometry - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Flow Cytometry - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryo) cells labelling CDK9 with ab239364 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Flow Cytometry - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Flow Cytometry - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma) cells labelling CDK9 with ab239364 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • ChIP - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    ChIP - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

    Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab239364 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G Sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). 
    Primers and probes are from paper PMCID: PMC2756882.

    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239364).

  • Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)
    Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade – BSA and Azide free (ab259268)

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