Human CDK4 knockout HeLa cell lysate
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CDK4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 139 bp insertion in exon2 and 2 bp deletion in exon2.
View Alternative Names
CDK4 protein, CDK4_HUMAN, CMM 3, Cell division kinase 4, Cell division protein kinase 4, Crk3, Cyclin-dependent kinase 4, MGC14458, Melanoma cutaneous malignant 3, PSK-J3, p34 cdk4
- WB
Lab
Western blot - Human CDK4 knockout HeLa cell lysate (AB263780)
Lane 1 : Wild-type HeLa cell lysate (20µg)
Lane 2 : CDK4 knockout HeLa cell lysate (20µg)
Lanes 1- 2 : Merged signal (red and green). Green - ab108357 observed at 34 kDa. Red - loading control ab130007 observed at 124 kDa.
ab108357 Recombinant Anti-Cdk4 antibody [EPR4513-32-7] was shown to specifically react with CDK4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255378 (knockout cell lysate ab263780) was used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108357 and Anti-Vinculin antibody [VIN-54] were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (<a href='/en-us/products/primary-antibodies/cdk4-antibody-epr4513-32-7-ab108357'>ab108357</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDK4 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDK4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cdk4-knockout-hela-cell-line-ab255378'>ab255378</a>)
Predicted band size: 34 kDa,55 kDa
Observed band size: 34 kDa,55 kDa
false
- WB
Lab
Western blot - Human CDK4 knockout HeLa cell lysate (AB263780)
Lane 1 : Wild-type HeLa cell lysate (20µg)
Lane 2 : CDK4 knockout HeLa cell lysate (20µg)
Lanes 1- 2 : Merged signal (red and green). Green - ab199728 observed at 34 kDa. Red - loading control ab130007 observed at 124 kDa.
ab199728 Recombinant Anti-Cdk4 antibody [EPR17525] was shown to specifically react with CDK4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255378 (knockout cell lysate ab263780) was used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab199728 and Anti-Vinculin antibody [VIN-54] were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Cdk4 antibody [EPR17525] (<a href='/en-us/products/primary-antibodies/cdk4-antibody-epr17525-ab199728'>ab199728</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDK4 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDK4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cdk4-knockout-hela-cell-line-ab255378'>ab255378</a>)
Predicted band size: 34 kDa
Observed band size: 34 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CDK4 knockout HeLa cell lysate (AB263780)
Allele-1 : 2 bp deletion in exon2; Allele-2 : 139 bp insertion in exon2
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Cdk4 plays a fundamental role in cell cycle regulation by forming a complex with D-type cyclins. Together they phosphorylate the retinoblastoma protein (Rb) resulting in the release of E2F transcription factors which promote the expression of genes necessary for DNA replication. The Cdk4/cyclin D complex regulates the cell's commitment to enter S phase and continue cell division. This regulation is key for normal cell proliferation and tissue homeostasis. Cdk4 activity is strictly controlled by INK4 family members acting as inhibitors and adding an extra regulation layer.
Pathways
Cdk4 is an integral part of important signaling pathways like the cell cycle and PI3K/AKT pathways. In the cell cycle pathway Cdk4 relays signals downstream that drive the transition from the G1 to S phase through its interaction with cyclin D and Rb. In the PI3K/AKT pathway signaling can influence cyclin D expression indirectly affecting Cdk4 activity. Proteins such as Cdk6 closely relate to Cdk4 and often compensate or partner with Cdk4 in these pathways to maintain proper cell cycle progression.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB199728
20ul selling size|RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR17525] (AB199728)](https://content.abcam.com/products/images/cdk4-antibody-epr17525-ab199728--immunocytochemistry-immunofluorescence-img77408.jpg)
- 0
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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