Human PKN2 knockout HeLa cell lysate (ab258587)
Overview
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Product name
Human PKN2 knockout HeLa cell lysate
See all PKN2 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5. -
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 ab260589 - Human PKN2 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin 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
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
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260589 - Human PKN2 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin 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
Images
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Western blot - Human PKN2 knockout HeLa cell lysate (ab258587)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: PKN2 knockout HeLa cell lysate (20 µg)
Lane 3: 293T cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab138514 observed at 112 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab138514 Anti-PKN2 antibody [EPR5490] was shown to specifically react with PKN2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264691 (knockout cell lysate ab258587) 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 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 10000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human PKN2 knockout HeLa cell lysate (ab258587)Homozygous: 1 bp insertion in exon 5
