Invitrogen

CDK6 Recombinant Rabbit Monoclonal Antibody (SD20-50)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

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Datasheet

Questions & Answers

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CDK6 Antibody in Immunocytochemistry (ICC) — Immunofluorescence analysis of CDK6 was performed using 70% confluent log phase MOLT-4 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CDK6 Recombinant Rabbit Monoclonal Antibody (SD20-50) (Product # MA5-32357) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

CDK6 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of CDK6 of paraffin-embedded Human colon cancer tissue using a CDK6 Monoclonal antibody (Product #MA5-32357). Counter stained with hematoxylin.

CDK6 Antibody in Western Blot (WB) — Western blot was performed using Anti-CDK6 Recombinant Rabbit Monoclonal Antibody (SD20-50) (Product # MA5-32357) and a 38 kDa band corresponding to CDK6 was observed in Jurkat, MOLT-4 but not in HeLa and MCF7. Whole cell extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), HeLa (Lane 3) and MCF7 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution), using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

CDK6 Antibody in Flow Cytometry (Flow) — Flow Cytometric analysis of CDK6 in Hela cells using a CDK6 Monoclonal Antibody (Product # MA5-32357) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.

CDK6 Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines owing to their inherent genetic constitution. The expression was observed in Jurkat and MOLT-4 and not in HeLa and MCF7 using CDK6 Recombinant Rabbit Monoclonal Antibody (SD20-50) (Product # MA5-32357) in Western Blot. {RE}

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1:50-1:100

Immunocytochemistry (ICC)

1:50-1:200

Immunofluorescence (IF)

1:50-1:200

Immunohistochemistry (IHC)

1:50-1:200

Western Blot (WB)

1:1,000-1:2,000

Product Specifications

Species

Human

Expression System

Rabbit / IgG

Class

Recombinant Monoclonal

Type

Antibody

Clone

SD20-50

Immunogen

Synthetic peptide within C-terminal human Cdk6

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 40% Glycerol, 0.05% BSA

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2809638

Product Specific Information

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

Cyclin-dependent kinase 6 (p40cdk6 also known as PLSTIRE) belongs to a family of proteins designated as cdks. They are critical regulators of cell cycle progression. The prototype member of this family, p34cdc2, and a related protein cdk2, function late in the cycle while cdk4 and cdk6 are critically involved in G1 to S progression. CDK6 is essential for cell proliferation within the dentate gyrus of the hippocampus and the cuventricular zone of the lateral ventricles. Mutations in the gene results in microcephaly 12, primary, autosomal recessive (MCPH12).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.