Human PGAM5 knockout HeLa cell lysate (ab257581)
Overview
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Product name
Human PGAM5 knockout HeLa cell lysate
See all PGAM5 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1. -
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 ab262124 - Human PGAM5 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
Displays phosphatase activity for serine/threonine residues, and, dephosphorylates and activates MAP3K5 kinase. Has apparently no phosphoglycerate mutase activity. May be regulator of mitochondrial dynamics. Substrate for a KEAP1-dependent ubiquitin ligase complex. Contributes to the repression of NFE2L2-dependent gene expression. -
Sequence similarities
Belongs to the phosphoglycerate mutase family. BPG-dependent PGAM subfamily. -
Domain
According to PubMed:18387606, may contain a non-cleaved N-terminal mitochondrial targeting sequence that targets PGAM5 to the cytosolic side of the outer mitochondrial membrane, instead of a N-terminal transmembrane. -
Cellular localization
Mitochondrion. Mitochondrion outer membrane. Membrane. Isoform 2 overexpression results in the formation of disconnected punctuate mitochondria distributed throughout the cytoplasm. Isoform 1 overexpression results in the clustering of mitochondria around the nucleus. - Information by UniProt
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Alternative names
- Bcl-XL-binding protein v68
- BXLBv68
- MGC5352
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262124 - Human PGAM5 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 PGAM5 knockout HeLa cell lysate (ab257581)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: PGAM5 knockout HeLa cell lysate (20 µg)
Lane 3: HepG2 cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab244218 observed at 32 kDa. Red - loading control ab52866 observed at 50 kDa.
ab244218 Anti-PGAM5 antibody [CL0624] was shown to specifically react with PGAM5 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265141 (knockout cell lysate ab257581) was used. Wild-type and PGAM5 knockout samples were subjected to SDS-PAGE. ab244218 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 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.
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Sanger Sequencing - Human PGAM5 knockout HeLa cell lysate (ab257581)Homozygous: 1 bp insertion in exon1