CDK6 Antibody - #DF6448

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Product: CDK6 Antibody
Catalog: DF6448
Description: Rabbit polyclonal antibody to CDK6
Application: WB IHC
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken
Mol.Wt.: 36kDa; 37kD(Calculated).
Uniprot: Q00534
RRID: AB_2838411

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 200ul $350 In stock

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

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(88%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
CDK6 Antibody detects endogenous levels of total CDK6.
RRID:
AB_2838411
Cite Format: Affinity Biosciences Cat# DF6448, RRID:AB_2838411.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

CDK 6; CDK6; CDK6_HUMAN; Cell division protein kinase 6; Crk 2; Crk2; Cyclin dependent kinase 6; Cyclin-dependent kinase 6; MCPH12; MGC59692; p40; PLSTIRE; Serine/threonine protein kinase PLSTIRE; Serine/threonine-protein kinase PLSTIRE; STQTL11;

Immunogens

Immunogen:

A synthesized peptide derived from human CDK6, corresponding to a region within N-terminal amino acids.

Uniprot:

Q00534(CDK6_HUMAN) >>Visit HPA database.

Gene(ID):
CDK6(1021)
Expression:
Q00534 CDK6_HUMAN:

Expressed ubiquitously. Accumulates in squamous cell carcinomas, proliferating hematopoietic progenitor cells, beta-cells of pancreatic islets of Langerhans, and neuroblastomas. Reduced levels in differentiating cells.

Description:
The cyclin-dependent kinases form complexes with their cyclin partners and with CDK inhibitors. CDK6 and CDK4 associate with the D-type cyclins and target the retinoblastoma protein, allowing passage through the G1/S phase restriction point (1). CDK6/cyclin D complexes are sequestered in their inactive form through binding to one of the INK4 CDK inhibitor family members (2,3). Unlike the INK4 family of cdk inhibitors, the CDK inhibitor p21 Waf1/Cip1 may enhance the association of CDK4 and CDK6 with their cyclin D partners (4).
Sequence:
MEKDGLCRADQQYECVAEIGEGAYGKVFKARDLKNGGRFVALKRVRVQTGEEGMPLSTIREVAVLRHLETFEHPNVVRLFDVCTVSRTDRETKLTLVFEHVDQDLTTYLDKVPEPGVPTETIKDMMFQLLRGLDFLHSHRVVHRDLKPQNILVTSSGQIKLADFGLARIYSFQMALTSVVVTLWYRAPEVLLQSSYATPVDLWSVGCIFAEMFRRKPLFRGSSDVDQLGKILDVIGLPGEEDWPRDVALPRQAFHSKSAQPIEKFVTDIDELGKDLLLKCLTFNPAKRISAYSALSHPYFQDLERCKENLDSHLPPSQNTSELNTA

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
Dog
100
Chicken
100
Rabbit
100
Zebrafish
88
Xenopus
75
Sheep
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

Serine/threonine-protein kinase involved in the control of the cell cycle and differentiation; promotes G1/S transition. Phosphorylates pRB/RB1 and NPM1. Interacts with D-type G1 cyclins during interphase at G1 to form a pRB/RB1 kinase and controls the entrance into the cell cycle. Involved in initiation and maintenance of cell cycle exit during cell differentiation; prevents cell proliferation and regulates negatively cell differentiation, but is required for the proliferation of specific cell types (e.g. erythroid and hematopoietic cells). Essential for cell proliferation within the dentate gyrus of the hippocampus and the subventricular zone of the lateral ventricles. Required during thymocyte development. Promotes the production of newborn neurons, probably by modulating G1 length. Promotes, at least in astrocytes, changes in patterns of gene expression, changes in the actin cytoskeleton including loss of stress fibers, and enhanced motility during cell differentiation. Prevents myeloid differentiation by interfering with RUNX1 and reducing its transcription transactivation activity, but promotes proliferation of normal myeloid progenitors. Delays senescence. Promotes the proliferation of beta-cells in pancreatic islets of Langerhans. May play a role in the centrosome organization during the cell cycle phases.

PTMs:

Thr-177 phosphorylation and Tyr-24 dephosphorylation promotes kinase activity.

Subcellular Location:

Cytoplasm. Nucleus. Cell projection>Ruffle. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome.
Note: Localized to the ruffling edge of spreading fibroblasts. Kinase activity only in nucleus. Localized to the cytosol of neurons and showed prominent staining around either side of the nucleus (By similarity). Present in the cytosol and in the nucleus in interphase cells and at the centrosome during mitosis from prophase to telophase (PubMed:23918663).

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

Expressed ubiquitously. Accumulates in squamous cell carcinomas, proliferating hematopoietic progenitor cells, beta-cells of pancreatic islets of Langerhans, and neuroblastomas. Reduced levels in differentiating cells.

Family&Domains:

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

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

References

1). miR-30d Attenuates Pulmonary Arterial Hypertension via Targeting MTDH and PDE5A and Modulates the Beneficial Effect of Sildenafil. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2024 (PubMed: 39206778) [IF=15.1]

Application: WB    Species: Rat    Sample:

Figure 1 miR-30d overexpression attenuates pulmonary hypertension and vascular remodeling. (A and B) Relative miR-30d expression level in plasma samples (A) and in plasma-derived extracellular vesicles (EVs) (B) from patients with idiopathic pulmonary arterial hypertension (PAH) versus healthy controls (n = 10 vs 8). C) Relative miR-30d expression level in lung tissues of rats with monocrotaline (MCT)-induced pulmonary hypertension (PH) (n = 10-13). (D and E) Right ventricular systolic pressure (RVSP) (D) and Fulton index (RV/(LV+S) ratio) (E) were assessed (n = 6-7). F) Immunohistochemical staining with α-SMA antibody for analysis of the number of muscularized distal pulmonary arteries (10–50 µm in diameter, scale bar = 20 µm) and the medial wall thickness of pulmonary arteries (50–150 µm in diameter, scale bar = 100 µm) (n = 5-6). G) qRT-PCR for CDK1, CDK6, and CCNA2 mRNA levels in lung tissues (n = 6). H) Western blot for CDK6 in lung tissues (n = 6). Data are shown as means ± SD. Data between 2 groups were compared by Mann-Whitney U test for (A) and by independent-sample two-tailed Student's t-test for (B) and (C). Data among 4 groups were compared by robust two-way ANOVA test followed by post-hoc pairwiseMedianTest using the rcompanion package for CCNA2 in (G), and by two-way ANOVA test followed by Tukey post hoc test for data in other figures. *P < 0.05; **P < 0.01; ***P < 0.001.
2). d-Borneol enhances cisplatin sensitivity via p21/p27-mediated S-phase arrest and cell apoptosis in non-small cell lung cancer cells and a murine xenograft model. Cellular & Molecular Biology Letters, 2022 (PubMed: 35883026) [IF=9.2]
3). miR-34a-5p functions as a tumor suppressor in head and neck squamous cell cancer progression by targeting Flotillin-2. International journal of biological sciences, 2021 (PubMed: 34803501) [IF=8.2]

Application: WB    Species: human    Sample: HN4 and CAL27 cells

Figure 4. Downregulation of FLOT-2 inhibited the proliferation, migratory, invasive activity and cell cycle in HN4 and CAL27 cells. A. Expression of FLOT-2 mRNAs in primary tumor and normal samples in the TCGA HNSCC database. B. Low and high expression of FLOT-2 protein in HNSCC tissues were detected by IHC. C. Kaplan-Meier analysis to plot the overall survival curves and disease-free survival curves of HNSCC patients with expression of FLOT-2 and different clinicopathological characteristics and statistical significance was assessed, and it is indicated that high expression of FLOT-2 was associated with the poor prognosis and disease-free survival. D and E. Real-time RT-PCR assay and western blotting analyses were performed to detect the expression of FLOT-2 at the mRNA and protein levels, in HOK, HN4, HN6 and CAL27 cells, respectively. F and G. Real-time RT-PCR and western blotting analyses were performed to measure the expression of FLOT-2 in HN4 and CAL27 cells after FLOT-2 siRNA transfection, respectively. H. HN4 and CAL27 were transfection with si-FLOT-2, and the proliferation ability of cells was detected by CCK8 assay. I and J. Migrative and invasive ability of HN4 and CAL27 cells transfected with si-FLOT-2 were analyzed using wound healing assay and transwell chamber assay. K. Cell cycle distribution was analyzed by flow cytometry. L and M. Western blot analysis was used to detect protein expression levels of EMT (L) and cell cycle regulators (M) markers in HN4 and CAL27 cells. Data were presented as the mean ± SEM from three independent experiments. *p < 0.05; **p < 0.01; ***p
4). Identification of RAC1 in promoting brain metastasis of lung adenocarcinoma using single-cell transcriptome sequencing. Cell death & disease, 2023 (PubMed: 37202394) [IF=8.1]

Application: WB    Species: human    Sample:

Fig. 5: RAC1 promotes the proliferation and cell cycle of tumor cells. a CCK-8 assay showed that RAC1 inhibitor could significantly slowed down the proliferation of A549 (Left panel) and H1975 (Right panel) cells at 48 h and 72 h (P 
5). Epigallocatechin gallate suppresses mitotic clonal expansion and adipogenic differentiation of preadipocytes through impeding JAK2/STAT3-mediated transcriptional cascades. Phytomedicine : international journal of phytotherapy and phytopharmacology, 2024 (PubMed: 38552377) [IF=6.7]
6). Alternative splicing of EZH2 regulated by SNRPB mediates hepatocellular carcinoma progression via BMP2 signaling pathway. iScience, 2024 (PubMed: 39850359) [IF=5.8]
7). SULF1 regulates malignant progression of colorectal cancer by modulating ARSH via FAK/PI3K/AKT/mTOR signaling. Cancer cell international, 2024 (PubMed: 38844922) [IF=5.8]
8). HMGB2 promotes chondrocyte proliferation under negative pressure through the phosphorylation of AKT. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2021 (PubMed: 34333060) [IF=4.6]
9). CircRNA circ_0072995 promotes the progression of epithelial ovarian cancer by modulating miR-147a/CDK6 axis. Aging-US, 2020 (PubMed: 32877369) [IF=3.9]

Application: WB    Species: Human    Sample: HO8910 and A2780 cells

Figure 5 CDK6 was directly targeted by miR-147a and was indirectly regulated by circ_0072995. (A) The target genes of miR-147a were predicted in databases (TargetScan, miRWalk, miRDB, StarBase) and there were 141 genes which overlapped in the Venn diagram. (B) PPI network analyses was performed by use of the String database, and results with a combined score of > 0.4 were selected. Among these, CDK6, AKT3, AKT2, PDPK1 were found to have had a high degree of connectivity and were selected as hub genes for miR-147a. (C) CDK6, AKT3, AKT2 expression was significantly reduced after overexpression of miR-147a, especially for CDK6. (D) There were multiple binding sites between miR-147a and CDK6. (E) The binding relationship between miR-147a and CDK6 was confirmed by dual-luciferase reporter assays. (F) miR-147a mimics markedly reduced expression of CDK6, whereas miR-147a inhibitors significantly enhanced CDK6 levels in HO8910 and A2780 cells. (G) Expression of CDK6 mRNA decreased after knockdown of circ_0072995, but was rescued by miR-147a inhibitors as determined by qRT-PCR. (H) The expression of CDK6 proteins decreased after knockdown of circ_0072995, but was rescued by miR-147a inhibitors as determined by WB.
10). miR-3619-5p inhibits prostate cancer cell growth by activating CDKN1A expression. ONCOLOGY REPORTS, 2017 (PubMed: 27878260) [IF=3.8]

Application: WB    Species: human    Sample:

(D) Expression of cyclin D1, CDK4 and CDK6 mRNA was determined by western blot analysis. α-Tublin served as a loading control. *
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