CDK4 Monoclonal antibody

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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.

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  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.

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  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.

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  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).

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  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).

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

Cat No. 66950-1-Ig

Clone No.1G2C12

Publications(31)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, FC (Intra), ELISA

Synonyms

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

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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 30 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
• UNIPROT ID: P11802
• 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