CDK4 Monoclonal antibody

CDK4 Monoclonal Antibody for WB, IHC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

28

CloneNo.

1G2C12

Cat no : 66950-1-Ig

Synonyms

PSK J3, EC:2.7.11.22, Cyclin-dependent kinase 4, cyclin dependent kinase 4, CMM3

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Validation Data Gallery

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  WB analysis using 66950-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

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  WB analysis of 4T1 using 66950-1-Ig

  4T1 cells were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66950-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 66950-1-Ig

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 66950-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 66950-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66950-1-Ig (CDK4 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 66950-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66950-1-Ig (CDK4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66950-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66950-1-Ig (CDK4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66950-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66950-1-Ig (CDK4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 66950-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66950-1-Ig (CDK4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  FC experiment of MCF-7 using 66950-1-Ig

  1X10^6 MCF-7 cells were intracellularly stained with 0.2 ug Anti-Human CDK4 (66950-1-Ig, Clone:1G2C12) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: LNCaP cells, HeLa cells, HEK-293 cells, 4T1 cells, Jurkat cells, MCF-7 cells, HepG2 cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, RAW 264.7 cells
• Positive IHC detected in: human breast cancer tissue, human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC (Intra) detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:200-1:1000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 28 publications below
• IHC: See 1 publications below

Product Information

66950-1-Ig targets CDK4 in WB, IHC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: CDK4 fusion protein Ag20538
• Full Name: cyclin-dependent kinase 4
• Calculated Molecular Weight: 34 kDa
• Observed Molecular Weight: 34 kDa
• GenBank Accession Number: BC010153
• Gene Symbol: CDK4
• Gene ID (NCBI): 1019
• RRID: AB_2882273
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Cyclin-dependent kinase-4 (CDK4) is a protein-serine kinase involved in the cell cycle. It is essential for the G1- to S-phase transition during the cell cycle and its expression is primarily controlled at the transcriptional level(PMID:17253961). CCND1-CDK4 axis is not only critical in glial tumor cells but also in stromal-derived cells in the surrounding tumor microenvironment that are vital to sustain tumor outgrowth(PMID:21844184).

Publications

Species	Application	Title
human	WB	Phytomedicine; Transcriptomics and molecular docking reveal the potential mechanism of lycorine against pancreatic cancer; Authors - Xin Zhou
human	WB	J Med Chem; Design, Synthesis, and Biological Evaluation for First GPX4 and CDK Dual Inhibitors; Authors - Jiangmin Zhu
human	WB	Cell Oncol (Dordr); LINC02154 promotes the proliferation and metastasis of hepatocellular carcinoma by enhancing SPC24 promoter activity and activating the PI3K-AKT signaling pathway.; Authors - Huan Yue
mouse	WB	Front Cell Dev Biol; LHPP-Mediated Histidine Dephosphorylation Suppresses the Self-Renewal of Mouse Embryonic Stem Cells.; Authors - Rong Mu Xia
human	WB	Front Cell Dev Biol; NLGN3 Upregulates Expression of ADAM10 to Promote the Cleavage of NLGN3 via Activating the LYN Pathway in Human Gliomas.; Authors - Ning-Ning Dang
human	WB	Oxid Med Cell Longev; MICAL2 Contributes to Gastric Cancer Cell Proliferation by Promoting YAP Dephosphorylation and Nuclear Translocation; Authors - Chenxiang Qi