Product nameHuman Plcb1 (Phospholipase C beta 1) knockout HeLa cell lysate
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon5.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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 - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: After reconstitution, store the lysate at -80°C.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
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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.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab262160 - Human PLCB1 knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255929 - Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
FunctionThe production of the second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) is mediated by activated phosphatidylinositol-specific phospholipase C enzymes.
Involvement in diseaseEpileptic encephalopathy, early infantile, 12
Sequence similaritiesContains 1 C2 domain.
Contains 1 PI-PLC X-box domain.
Contains 1 PI-PLC Y-box domain.
Cellular localizationNucleus membrane. Cytoplasm. Colocalizes with the adrenergic receptors, ADREN1A and ADREN1B, at the nuclear membrane of cardiac myocytes.
- Information by UniProt
- 1 phosphatidylinositol 4,5 bisphosphate phosphodiesterase beta 1
- 1-phosphatidylinositol 4
KO cell lines
Our Abpromise guarantee covers the use of ab257589 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 139 kDa.|
Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PLCB1 knockout HeLa cell lysate (20 ug)
Lane 3:HepG2 cell lysate (20 ug)
Lane 4:Mouse heart tissue lysate (20 ug)
ab182359 was shown to specifically react with Phospholipase C beta 1/PLCB1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266033 (knockout cell lysate ab257589) was used. Wild-type and Phospholipase C beta 1/PLCB1 knockout samples were subjected to SDS-PAGE. ab182359 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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.
Homozygous: 1 bp insertion in exon5
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab257589 has not yet been referenced specifically in any publications.