Anti-Cyclin-dependent kinase 4 Cdk4 Antibody

Product Name

Anti-Cyclin-dependent kinase 4 Cdk4 Antibody

View all CDK4 Antibodies

SKU/Catalog Number

PA1428-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Cyclin-dependent kinase 4 Cdk4 Antibody catalog # PA1428-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Cyclin-dependent kinase 4 Cdk4 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1428-1)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of mouse Cdk4, different from the related rat sequence by one amino acid, and from the related human sequence by four amino acids.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1428-1 is reactive to CDK4 in Human, Mouse, Rat

Applications

PA1428-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

34 kDa

Calculated molecular weight

33.73kDa

Background of CDK4

Cyclin-dependent kinase-4 (CDK4) is a protein-serine kinase involved in the cell cycle. Human cell division is regulated primarily at the G1-to-S or the G2-to-M boundaries within the cell cycle. The complexes formed by CDK4 and the D-type cyclins are involved in the control of cell proliferation during the G1 phase. CDK4 is inhibited by p16, also known as cyclin-dependent kinase inhibitor-2. CDK4 is mapped to 12q14. CDK4 expression and activity are required for cytokine responsiveness in T cells.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CDK4 using anti-CDK4 antibody (PA1428-1, Left) and anti-CDK4 antibody (PB9535, Right).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: rat PC-12 whole cell lysates,
Lane 4: mosue NIH/3T3 whole cell lysates,
Lane 5: mosue RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CDK4 antigen affinity purified polyclonal antibody (Catalog # PA1428-1) and rabbit anti-CDK4 antigen affinity purified polyclonal antibody (Catalog # PB9535)at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CDK4 at approximately 34 kDa. The expected band size for CDK4 is at 34 kDa.

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Figure 2. IF analysis of CDK4 using anti-CDK4 antibody (PA1428-1) and anti-Tubulin Alpha antibody (M03989-3).
CDK4 was detected in immunocytochemical section of MCF-7 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-CDK4 Antibody (PA1428-1) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of MCF-7 cells using anti-CDK4 antibody (PA1428-1).
Overlay histogram showing MCF-7 cells stained with PA1428-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CDK4 Antibody (PA1428-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For CDK4 (Source: Uniprot.org, NCBI)

Alternative Names

CDK4; Cell division protein kinase 4; CMM3; cyclin-dependent kinase 4; EC 2.7.11; EC 2.7.11.22; melanoma cutaneous malignant, 3; MGC14458; PSK-J3; PSK-J3cell division kinase 4 CDK4 CMM3, PSK-J3 cyclin dependent kinase 4 cyclin-dependent kinase 4|cell division protein kinase 4

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