Human CDK6 knockout HeLa cell lysate
Be the first to review this product! Submit a review
|
(0 Publication)
CDK6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
View Alternative Names
CDK6_HUMAN, Cell division protein kinase 6, Crk 2, Cyclin-dependent kinase 6, MCPH12, MGC59692, PLSTIRE, STQTL11, Serine/threonine-protein kinase PLSTIRE
- WB
Lab
Western blot - Human CDK6 knockout HeLa cell lysate (AB257088)
Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : CDK6 knockout HeLa cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab241554 observed at 38 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab241554 was shown to react with Cdk6 in wild-type HeLa cells in Western blot with loss of signal observed in CDK6 knockout cell line ab266059 (CDK6 knockout cell lysate ab257088). Wild-type HeLa and CDK6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab241554 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-Cdk6 antibody [98D] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-98d-ab241554'>ab241554</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDK6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDK6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cdk6-knockout-hela-cell-line-ab266059'>ab266059</a>)
Predicted band size: 37 kDa
Observed band size: 38 kDa
false
- WB
Lab
Western blot - Human CDK6 knockout HeLa cell lysate (AB257088)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : CDK6 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab124821 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab124821 Anti-Cdk6 antibody [EPR4515] was shown to specifically react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type and Cdk6 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. ab124821 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-Cdk6 antibody [EPR4515] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-epr4515-ab124821'>ab124821</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDK6 knockout HeLa cell lysate at 20 µg
Predicted band size: 37 kDa
Observed band size: 37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (AB257088)
Allele-1 : 1 bp insertion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (AB257088)
Allele-2 : Insertion of the selection cassette in exon2
- WB
Lab
Western blot - Human CDK6 knockout HeLa cell lysate (AB257088)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : CDK6 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab54576 observed at 37 kDa. Red - loading control ab52866 observed at 50 kDa.
ab54576 Anti-Cdk6 antibody was shown to specifically react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type and Cdk6 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. ab54576 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Cdk6 antibody [8H4] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-8h4-ab54576'>ab54576</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDK6 knockout HeLa cell lysate (ab257088) at 20 µg
Predicted band size: 37 kDa
Observed band size: 37 kDa
false
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
Cdk6 functions as a regulator of the cell cycle. It participates in a complex with cyclin D to control the G1 phase progression. This activity is essential for the regulation of cell division and proliferation. The association with cyclin D allows Cdk6 to phosphorylate the retinoblastoma protein (Rb) leading to the release of E2F transcription factors that promote the expression of genes necessary for DNA synthesis.
Pathways
Cdk6 is an important component of the cell cycle control pathway. It interacts primarily with the retinoblastoma protein and Cyclin D1 in this context. The cell cycle pathway is critical for controlled cell proliferation and its dysregulation can lead to diseases like cancer. Another involved pathway is the PI3K/AKT pathway which can activate Cdk6 activity through upstream signaling events implicating Cdk6 in cellular responses to growth signals.
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
![Western blot - Anti-Cdk6 antibody [98D] (AB241554)](https://content.abcam.com/products/images/cdk6-antibody-98d-ab241554--western-blot-img53243.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com