Immunofluorescence (IF)
(Figure 2. IF analysis of CDK12 using anti-CDK12 antibody (MBS1753598).CDK12 was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- CDK12 Antibody (MBS1753598) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Rabbit anti-Human CDK12 Polyclonal Antibody | anti-CDK12 antibody
Anti-CDK12 Antibody
ICC/IF: Concentration: 5ug/ml; Tested Sepcies: Human
FC: Concentration: 1-3ug/1x106 cells; Tested Sepcies: Human
Direct ELISA: Concentration: 0.1-0.5ug/ml; Tested Sepcies: Human
Tested Species: In-house tested with positive results
Other applications have not been tested.
Optimal dilutions should be determined by end users.
Immunofluorescence (IF)
(Figure 2. IF analysis of CDK12 using anti-CDK12 antibody (MBS1753598).CDK12 was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- CDK12 Antibody (MBS1753598) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of CDK12 using anti-CDK12 antibody (MBS1753598).CDK12 was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- CDK12 Antibody (MBS1753598) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of HL-60 cells using anti-CDK12 antibody (MBS1753598).Overlay histogram showing HL-60 cells stained with MBS1753598 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CDK12 Antibody (MBS1753598, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of HL-60 cells using anti-CDK12 antibody (MBS1753598).Overlay histogram showing HL-60 cells stained with MBS1753598 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CDK12 Antibody (MBS1753598, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )
Western Blot (WB)
(Figure 1. Western blot analysis of CDK12 using anti-CDK12 antibody (MBS1753598).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 lysatesLane 2: human HELA whole cell lysatesLane 3: human Jurkat whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human HL-60 whole cell lysatesLane 6: human CACO-2 whole cell lysatesLane 7: human A549 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 rabbit anti-CDK12 antigen affinity purified polyclonal antibody (Catalog # MBS1753598) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for CDK12 at approximately 205KD. The expected band size for CDK12 is at 205KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of CDK12 using anti-CDK12 antibody (MBS1753598).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 lysatesLane 2: human HELA whole cell lysatesLane 3: human Jurkat whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human HL-60 whole cell lysatesLane 6: human CACO-2 whole cell lysatesLane 7: human A549 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 rabbit anti-CDK12 antigen affinity purified polyclonal antibody (Catalog # MBS1753598) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for CDK12 at approximately 205KD. The expected band size for CDK12 is at 205KD. )
2. Cheng, S. -W. G., Kuzyk, M. A., Moradian, A., Ichu, T. -A., Chang, V. C. -D., Tien, J. F., Vollett, S. E., Griffith, M., Marra, M. A., Morin, G. B. Interaction of cyclin-dependent kinase 12/CrkRS with cyclin K1 is required for the phosphorylation of the C-terminal domain of RNA polymerase II. Molec. Cell. Biol. 32: 4691-4704, 2012.
3. Dubbury, S. J., Boutz, P. L., Sharp, P. A. CDK12 regulates DNA repair genes by suppressing intronic polyadenylation. Nature 564: 141-145, 2018.
NCBI and Uniprot Product Information
Uniprot Description
CDK12: a Ser/Thr protein kinase of the CMGC group and CDK family. Phosphorylates the C-terminal heptapeptide repeat domain (CTD) of the largest RNA polymerase II subunit RPB1, thereby acting as a key regulator of transcription elongation. Required for RNA splicing, possibly by phosphorylating SRSF1/SF2. Interacts with CCNL1 and CCNL2. Colocalized with nuclear speckles throughout interphase. Has a highly phosphorylated arginine/serine-rich (RS) domain N-terminal to the kinase domain. Chromosomal aberrations involving CDK12 may be a cause gastric cancer. Deletions within 17q12 region producing fusion transcripts with ERBB2, leading to CDK12-ERBB2 fusion leading to trunctated CDK12 protein not in-frame with ERBB2. Three isoforms of the human protein are produced by alternative splicing.
Protein type: EC 2.7.11.22; Motility/polarity/chemotaxis; Protein kinase, Ser/Thr (non-receptor); Kinase, protein; Cell cycle regulation; Protein kinase, CMGC; RNA splicing; EC 2.7.11.23; Spliceosome; CMGC group; CDK family; CRK7 subfamily; CDK/CRK7 subfamily
Chromosomal Location of Human Ortholog: 17q12
Cellular Component: nucleoplasm; nuclear cyclin-dependent protein kinase holoenzyme complex; nucleolus; nuclear speck; nucleus
Molecular Function: RNA polymerase subunit kinase activity; protein binding; cyclin binding; cyclin-dependent protein kinase activity; ATP binding; protein kinase activity
Biological Process: regulation of cell cycle; protein amino acid autophosphorylation; RNA splicing; regulation of MAP kinase activity; mRNA processing
Research Articles on CDK12
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Product Notes
The CDK12 cdk12 (Catalog #AAA1753598) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CDK12 Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's CDK12 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: Concentration: 0.25-0.5ug/ml; Tested Species: Human ICC/IF: Concentration: 5ug/ml; Tested Sepcies: Human FC: Concentration: 1-3ug/1x106 cells; Tested Sepcies: Human Direct ELISA: Concentration: 0.1-0.5ug/ml; Tested Sepcies: Human Tested Species: In-house tested with positive results Other applications have not been tested. Optimal dilutions should be determined by end users. Researchers should empirically determine the suitability of the CDK12 cdk12 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CDK12, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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