Human BBC3 (PUMA) knockout HeLa cell lysate
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(0 Publication)
- WB
Lab
Western blot - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)
Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : BBC3 knockout HeLa cell lysate 20 ug
Lane 3 : A549 cell lysate 20 ug
Lane 4 : PC-3 cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - anti-Puma antibody observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
anti-Puma antibody was shown to react with Puma in wild-type HeLa cells in Western blot with loss of signal observed in BBC3 knockout cell line ab261832 (BBC3 knockout cell lysate ab256848). Wild-type HeLa and BBC3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with anti-Puma antibody and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Anti-Puma antibody at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
BBC3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human BBC3 (PUMA) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-bbc3-puma-knockout-hela-cell-line-ab261832'>ab261832</a>)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
PC-3 cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)
Allele-1 : 22 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)
Allele-2 : 10 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)
Allele-3 : 1 bp insertion in exon2
Complementary Products
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
PUMA functions as a pivotal mediator of apoptosis in response to diverse stimuli such as DNA damage and oncogenic stress. As part of the Bcl-2 family PUMA does not typically form a complex with other proteins but functions directly by binding and neutralizing anti-apoptotic Bcl-2 family proteins. This action results in the release of caspase-activating factors from mitochondria essential for programmed cell death.
Pathways
Several cellular pathways incorporate PUMA to regulate apoptosis. PUMA is notably included in the p53 pathway where p53 transcriptionally activates PUMA following cellular stress or DNA damage. The presence of related proteins like Bax and Bim in the mitochondrial apoptotic pathway links these signals to the mitochondrial death machinery facilitating apoptosis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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