Product: Phospho-CDK9 (Thr186) Antibody
Catalog: AF4440
Description: Rabbit polyclonal antibody to Phospho-CDK9 (Thr186)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Dog, Chicken
Mol.Wt.: 42,55kDa; 43kD(Calculated).
Uniprot: P50750
RRID: AB_2844504

View similar products>>

   Size Price Inventory
 100ul $350 In stock
 200ul $450 In stock

Lead Time: Same day delivery

For pricing and ordering contact:
Local distributors

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(92%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
Phospho-CDK9 (Thr186) Antibody detects endogenous levels of CDK9 only when phosphorylated at Thr186.
RRID:
AB_2844504
Cite Format: Affinity Biosciences Cat# AF4440, RRID:AB_2844504.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

C-2K; CDC2 related kinase; CDC2L4; Cdk 9; Cdk9; CDK9_HUMAN; Cell division cycle 2-like protein kinase 4; Cell division protein kinase 9; CTK1; Cyclin dependent kinase 9; Cyclin-dependent kinase 9; PITALRE; Serine/threonine-protein kinase PITALRE; TAK; Tat associated kinase complex catalytic subunit;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P50750 CDK9_HUMAN:

Ubiquitous.

Sequence:
MAKQYDSVECPFCDEVSKYEKLAKIGQGTFGEVFKARHRKTGQKVALKKVLMENEKEGFPITALREIKILQLLKHENVVNLIEICRTKASPYNRCKGSIYLVFDFCEHDLAGLLSNVLVKFTLSEIKRVMQMLLNGLYYIHRNKILHRDMKAANVLITRDGVLKLADFGLARAFSLAKNSQPNRYTNRVVTLWYRPPELLLGERDYGPPIDLWGAGCIMAEMWTRSPIMQGNTEQHQLALISQLCGSITPEVWPNVDNYELYEKLELVKGQKRKVKDRLKAYVRDPYALDLIDKLLVLDPAQRIDSDDALNHDFFWSDPMPSDLKGMLSTHLTSMFEYLAPPRRKGSQITQQSTNQSRNPATTNQTEFERVF

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Chicken
100
Zebrafish
92
Xenopus
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P50750 As Substrate

Site PTM Type Enzyme
K3 Acetylation
K3 Ubiquitination
S7 Phosphorylation
K18 Ubiquitination
K21 Methylation
K21 Ubiquitination
K24 Sumoylation
K24 Ubiquitination
T29 Phosphorylation
K35 Acetylation
K35 Methylation
K35 Ubiquitination
K44 Acetylation
K44 Ubiquitination
K48 Acetylation
K49 Acetylation
K49 Sumoylation
K49 Ubiquitination
K56 Acetylation
K56 Ubiquitination
K68 Acetylation
K68 Ubiquitination
K74 Acetylation
R86 Methylation
K88 Ubiquitination
S90 Phosphorylation P24941 (CDK2)
K127 Acetylation
K127 Methylation
K151 Sumoylation
K151 Ubiquitination
K164 Acetylation
K164 Ubiquitination
S175 Phosphorylation
K178 Acetylation
K178 Ubiquitination
S180 Phosphorylation
T186 Phosphorylation P50750 (CDK9)
K269 Acetylation
K269 Ubiquitination
K274 Acetylation
K276 Acetylation
K280 Ubiquitination
K294 Ubiquitination
S317 Phosphorylation
S329 Phosphorylation
T330 Phosphorylation
T333 Phosphorylation
S334 Phosphorylation
Y338 Phosphorylation
K345 Acetylation
K345 Sumoylation
K345 Ubiquitination
S347 Phosphorylation P50750 (CDK9)
T350 Phosphorylation
S353 Phosphorylation P50750 (CDK9)
T354 Phosphorylation P50750 (CDK9)
S357 Phosphorylation
T362 Phosphorylation P50750 (CDK9)
T363 Phosphorylation P50750 (CDK9)
T366 Phosphorylation
R370 Methylation

PTMs - P50750 As Enzyme

Substrate Site Source
O00267 (SUPT5H) S666 Uniprot
O00267 (SUPT5H) T768 Uniprot
O00267 (SUPT5H) T775 Uniprot
O00267 (SUPT5H) T784 Uniprot
O00267 (SUPT5H) T791 Uniprot
O00267 (SUPT5H) T799 Uniprot
O00267 (SUPT5H) T806 Uniprot
O00267 (SUPT5H) T814 Uniprot
P04637-1 (TP53) S33 Uniprot
P04637 (TP53) S315 Uniprot
P04637-1 (TP53) S392 Uniprot
P06400 (RB1) S795 Uniprot
P06400 (RB1) S807 Uniprot
P06400 (RB1) S811 Uniprot
P24928 (POLR2A) S1616 Uniprot
P24928 (POLR2A) T1618 Uniprot
P24928 (POLR2A) S1619 Uniprot
P24928 (POLR2A) S1621 Uniprot
P24928 (POLR2A) S1623 Uniprot
P24928 (POLR2A) S1644 Uniprot
P24928 (POLR2A) S1651 Uniprot
P24928 (POLR2A) S1665 Uniprot
P24928 (POLR2A) S1672 Uniprot
P24928 (POLR2A) S1693 Uniprot
P24928 (POLR2A) S1714 Uniprot
P24928 (POLR2A) S1721 Uniprot
P24928 (POLR2A) S1735 Uniprot
P24928 (POLR2A) S1763 Uniprot
P24928 (POLR2A) S1784 Uniprot
P24928 (POLR2A) S1861 Uniprot
P24928 (POLR2A) S1868 Uniprot
P24928 (POLR2A) S1875 Uniprot
P24928 (POLR2A) S1878 Uniprot
P24928 (POLR2A) S1882 Uniprot
P24928 (POLR2A) S1889 Uniprot
P24928 (POLR2A) S1896 Uniprot
P24928 (POLR2A) S1910 Uniprot
P24928 (POLR2A) S1917 Uniprot
P24928 (POLR2A) S1924 Uniprot
P24928 (POLR2A) S1931 Uniprot
P24928 (POLR2A) S1941 Uniprot
P24928 (POLR2A) S1948 Uniprot
P37231 (PPARG) S112 Uniprot
P50750 (CDK9) T186 Uniprot
P50750 (CDK9) S347 Uniprot
P50750 (CDK9) S353 Uniprot
P50750 (CDK9) T354 Uniprot
P50750 (CDK9) T362 Uniprot
P50750 (CDK9) T363 Uniprot
P84022 (SMAD3) T179 Uniprot
P84022 (SMAD3) S208 Uniprot
P84022 (SMAD3) S213 Uniprot
Q02539 (HIST1H1A) S183 Uniprot
Q15596 (NCOA2) S469 Uniprot
Q15596 (NCOA2) S487 Uniprot
Q15596 (NCOA2) S493 Uniprot
Q15596 (NCOA2) S499 Uniprot
Q15797 (SMAD1) S187 Uniprot
Q15797 (SMAD1) S195 Uniprot
Q15797 (SMAD1) S206 Uniprot
Q15797 (SMAD1) S214 Uniprot
Q6PGQ7 (BORA) S325 Uniprot
Q96PM5 (RCHY1) S211 Uniprot
Q96PM5 (RCHY1) T217 Uniprot
Q9H0D6 (XRN2) T439 Uniprot

Research Backgrounds

Function:

Protein kinase involved in the regulation of transcription. 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 productive elongation by phosphorylating the CTD (C-terminal domain) of the large subunit of RNA polymerase II (RNAP II) POLR2A, SUPT5H and RDBP. This complex is inactive when in the 7SK snRNP complex form. Phosphorylates EP300, MYOD1, RPB1/POLR2A and AR and the negative elongation factors DSIF and NELF. Regulates cytokine inducible transcription networks by facilitating promoter recognition of target transcription factors (e.g. TNF-inducible RELA/p65 activation and IL-6-inducible STAT3 signaling). Promotes RNA synthesis in genetic programs for cell growth, differentiation and viral pathogenesis. P-TEFb is also involved in cotranscriptional histone modification, mRNA processing and mRNA export. Modulates a complex network of chromatin modifications including histone H2B monoubiquitination (H2Bub1), H3 lysine 4 trimethylation (H3K4me3) and H3K36me3; integrates phosphorylation during transcription with chromatin modifications to control co-transcriptional histone mRNA processing. The CDK9/cyclin-K complex has also a kinase activity towards CTD of RNAP II and can substitute for CDK9/cyclin-T P-TEFb in vitro. Replication stress response protein; the CDK9/cyclin-K complex is required for genome integrity maintenance, by promoting cell cycle recovery from replication arrest and limiting single-stranded DNA amount in response to replication stress, thus reducing the breakdown of stalled replication forks and avoiding DNA damage. In addition, probable function in DNA repair of isoform 2 via interaction with KU70/XRCC6. Promotes cardiac myocyte enlargement. RPB1/POLR2A phosphorylation on 'Ser-2' in CTD activates transcription. AR phosphorylation modulates AR transcription factor promoter selectivity and cell growth. DSIF and NELF phosphorylation promotes transcription by inhibiting their negative effect. The phosphorylation of MYOD1 enhances its transcriptional activity and thus promotes muscle differentiation.

PTMs:

Autophosphorylation at Thr-186, Ser-347, Thr-350, Ser-353, Thr-354 and Ser-357 triggers kinase activity by promoting cyclin and substrate binding (e.g. HIV TAT) upon conformational changes. Thr-186 phosphorylation requires the calcium Ca(2+) signaling pathway, including CaMK1D and calmodulin. This inhibition is relieved by Thr-29 dephosphorylation. However, phosphorylation at Thr-29 is inhibitory within the HIV transcription initiation complex. Phosphorylation at Ser-175 inhibits kinase activity. Can be phosphorylated on either Thr-362 or Thr-363 but not on both simultaneously.

Dephosphorylation of Thr-186 by PPM1A and PPM1B blocks CDK9 activity and may lead to CDK9 proteasomal degradation. However, PPP1CA-mediated Thr-186 dephosphorylation is required to release P-TEFb from its inactive P-TEFb/7SK snRNP complex. Dephosphorylation of C-terminus Thr and Ser residues by protein phosphatase-1 (PP1) triggers CDK9 activity, contributing to the activation of HIV-1 transcription.

N6-acetylation of Lys-44 promotes kinase activity, whereas acetylation of both Lys-44 and Lys-48 mediated by PCAF/KAT2B and GCN5/KAT2A reduces kinase activity. The acetylated form associates with PML bodies in the nuclear matrix and with the transcriptionally silent HIV-1 genome; deacetylated upon transcription stimulation. Deacetylated by SIRT7, promoting the kinase activity and subsequent 'Ser-2' phosphorylation of the C-terminal domain (CTD) of RNA polymerase II.

Polyubiquitinated and thus activated by UBR5. This ubiquitination is promoted by TFIIS/TCEA1 and favors 'Ser-2' phosphorylation of RPB1/POLR2A CTD.

Subcellular Location:

Nucleus. Cytoplasm. Nucleus>PML body.
Note: Accumulates on chromatin in response to replication stress. Complexed with CCNT1 in nuclear speckles, but uncomplexed form in the cytoplasm. The translocation from nucleus to cytoplasm is XPO1/CRM1-dependent. Associates with PML body when acetylated.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Ubiquitous.

Subunit Structure:

Component of the super elongation complex (SEC), at least composed of EAF1, EAF2, CDK9, MLLT3/AF9, AFF (AFF1 or AFF4), the P-TEFb complex and ELL (ELL, ELL2 or ELL3). Associates with CCNT1/cyclin-T1, CCNT2/cyclin-T2 (isoform A and isoform B) or CCNK/cyclin-K to form active P-TEFb. P-TEFb forms a complex with AFF4/AF5Q31 and is part of the super elongation complex (SEC). Component of a complex which is composed of at least 5 members: HTATSF1/Tat-SF1, P-TEFb complex, RNA pol II, SUPT5H and NCL/nucleolin. Associates with UBR5 and forms a transcription regulatory complex composed of CDK9, RNAP II, UBR5 and TFIIS/TCEA1 that can stimulate target gene transcription (e.g. gamma fibrinogen/FGG) by recruiting their promoters. Component of the 7SK snRNP inactive complex which is composed of at least 8 members: P-TEFb (composed of CDK9 and CCNT1/cyclin-T1), HEXIM1, HEXIM2, LARP7, BCDIN3, SART3 proteins and 7SK and U6 snRNAs. This inactive 7SK snRNP complex can also interact with NCOR1 and HDAC3, probably to regulate CDK9 acetylation. Release of P-TEFb from P-TEFb/7SK snRNP complex requires both PP2B to transduce calcium Ca(2+) signaling in response to stimuli (e.g. UV or hexamethylene bisacetamide (HMBA)) and PPP1CA to dephosphorylate Thr-186. This released P-TEFb remains inactive in the pre-initiation complex with BRD4 until new Thr-186 phosphorylation occurs after the synthesis of a short RNA. Interacts with BRD4; to target chromatin binding. Interacts with JMJD6. Interacts with activated nuclear STAT3 and RELA/p65. Binds to AR and MYOD1. Forms a complex composed of CDK9, CCNT1/cyclin-T1, EP300 and GATA4 that stimulates hypertrophy in cardiomyocytes. The large PER complex involved in the repression of transcriptional termination is composed of at least PER2, CDK9, DDX5, DHX9, NCBP1 and POLR2A (By similarity). Interacts with HSF1. Interacts with TBX21 (By similarity). Isoform 3: binds to KU70/XRCC6. Interacts with WDR43 (By similarity).

(Microbial infection) Interacts with the acidic/proline-rich region of HIV-1 and HIV-2 Tat via T-loop region and is thus required for HIV to hijack host transcription machinery during its replication through cooperative binding to viral TAR RNA. Interacts with herpes simplex virus 1 protein ICP22; this interaction inhibits the positive transcription elongation factor b (P-TEFb).

Family&Domains:

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.

Research Fields

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

Restrictive clause

 

Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale. Not for distribution without written consent. Affinity Biosciences will not be held responsible for patent infringement or other violations that may occur with the use of our products. Affinity Biosciences, Affinity Biosciences Logo and all other trademarks are the property of Affinity Biosciences LTD.