Human PKN2 knockout HCT116 cell lysate (ab258588)
Overview
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Product name
Human PKN2 knockout HCT116 cell lysate
See all PKN2 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HCT116 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon5 and 23 bp deletion in exon5. -
Passage number
Knockout validation
Sanger Sequencing, Western Blot (WB)Reconstitution notes
To 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.
Notes
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: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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 applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263659 - Human PKN2 knockout HCT116 cell lysate 1 x 100µg ab255555 - Human wild-type HCT116 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X D5S818: 10, 11 D13S317: 10, 12 D7S820: 11, 12 D16S539: 11, 13 vWA: 17, 22 TH01: 8,9 TPOX: 8, 9 CSF1PO: 7, 10
Target
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Function
Exhibits a preference for highly basic protein substrates. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 C2 domain.
Contains 1 protein kinase domain.
Contains 3 REM (Hr1) repeats. -
Domain
The C1 domain does not bind the diacylglycerol (DAG). -
Post-translational
modificationsAutophosphorylated.
Activated by limited proteolysis with trypsin. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- Cardiolipin activated protein kinase Pak2
- MGC150606
- MGC71074
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263659 - Human PKN2 knockout HCT116 cell lysate 1 x 100µg ab255555 - Human wild-type HCT116 cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X D5S818: 10, 11 D13S317: 10, 12 D7S820: 11, 12 D16S539: 11, 13 vWA: 17, 22 TH01: 8,9 TPOX: 8, 9 CSF1PO: 7, 10
Images
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Western blot - Human PKN2 knockout HCT116 cell lysate (ab258588)
Lane 1:Wild-type HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 2:PKN2 knockout HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 3:HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)ab138514 was shown to specifically react with PKN2 in wild-type HCT cells. Loss of signal was observed when knockout cell line ab266894 (knockout cell lysate ab258588) was used. Wild-type and PKN2 knockout samples were subjected to SDS-PAGE. ab138514 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.
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Sanger Sequencing - Human PKN2 knockout HCT116 cell lysate (ab258588)Allele-1: 23 bp deletion in exon5
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Sanger Sequencing - Human PKN2 knockout HCT116 cell lysate (ab258588)Allele-2: 1 bp insertion in exon5