Recombinant Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade (ab239364)
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
- Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IF, ChIP, ChIC/CUT&RUN-seq, ChIP-sequencing
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade
See all Cdk9 primary antibodies -
Description
Rabbit monoclonal [EPR22956-37] to Cdk9 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IF, ChIP, ChIC/CUT&RUN-seq, ChIP-sequencingmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
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
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
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22956-37 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239364 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
1/600.
|
|
IP |
1/30.
|
|
WB |
1/1000. Detects a band of approximately 42, 55 kDa (predicted molecular weight: 43 kDa).
|
|
IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. |
|
ICC/IF |
1/100.
|
|
ChIP |
Use 5 µg for 25 µg of chromatin.
|
|
ChIC/CUT&RUN-seq |
Use 5µg for 105 cells.
|
|
ChIP-sequencing |
Use 8µg for 107 cells.
|
Notes |
---|
Flow Cyt (Intra)
1/600. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 42, 55 kDa (predicted molecular weight: 43 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. |
ICC/IF
1/100. |
ChIP
Use 5 µg for 25 µg of chromatin. |
ChIC/CUT&RUN-seq
Use 5µg for 105 cells. |
ChIP-sequencing
Use 8µg for 107 cells. |
Target
-
Function
Member of the cyclin-dependent kinase pair (CDK9/cyclin-T) complex, also called positive transcription elongation factor b (P-TEFb), which facilitates the transition from abortive to production elongation by phosphorylating the CTD (C-terminal domain) of the large subunit of RNA polymerase II (RNAP II), SUPT5H and RDBP. The CDK9/cyclin-K complex has also a kinase activity toward CTD of RNAP II and can substitute for P-TEFb in vitro. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. -
Cellular localization
Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 1025 Human
- Entrez Gene: 107951 Mouse
- Entrez Gene: 362110 Rat
- Omim: 603251 Human
- SwissProt: P50750 Human
- SwissProt: Q99J95 Mouse
- SwissProt: Q641Z4 Rat
- Unigene: 150423 Human
see all -
Alternative names
- C-2K antibody
- CDC2 related kinase antibody
- CDC2L4 antibody
see all
Images
-
CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 105 HeLa cells and 5µg of ab239364 [EPR22956-37]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 µg of ab239364 [EPR22956-37]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
-
Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8 µg of ab239364 [EPR22956-37]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
-
All lanes : Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade (ab239364) at 1/1000 dilution
Lane 1 : 293T (human embryonic kidney epithelial cell), whole cell lysate at 10 µg
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 3 : Mouse lung tissue lysate at 20 µg
Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg
Lane 5 : Mouse brain tissue lysate 20 ug
Lane 6 : Rat lung tissue lysate at 20 µg
Lane 7 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg
Lane 8 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg
Lane 9 : C6 (rat glial tumor glial cell) whole cell lysate at 10 µg
Secondary
Lanes 1-5 & 7-9 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 6 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 43 kDa
Observed band size: 42,55 kDa why is the actual band size different from the predicted?Blocking and Dilution Buffer and concentration: 5% NFDM/TBST
Exposure times.
Lanes 1-3: 36 seconds Lanes 4-5: 5.5 seconds Lane 6: 3 minutes Lanes 7-8: 15 seconds Lane 9: 48 seconds
-
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
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade (ab239364)
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.
-
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 (ab239364)
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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade (ab239364)
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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk9 antibody [EPR22956-37] - ChIP Grade (ab239364)
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.
-
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.
-
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.
-
Intracellular 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.
-
Intracellular 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.
-
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
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (0)
ab239364 has not yet been referenced specifically in any publications.