Human PTPN1 (PTP1B) knockout HeLa cell lysate (ab257617)
Overview
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Product name
Human PTPN1 (PTP1B) knockout HeLa cell lysate
See all PTP1B kits -
Product overview
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and Insertion of the selection cassette in exon 4. -
Passage number
Knockout validation
Sanger SequencingReconstitution 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 ab260304 - Human PTPN1 knockout HeLa cell lysate 1 x 100µg ab255552 - 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
May play an important role in CKII- and p60c-src-induced signal transduction cascades. -
Sequence similarities
Belongs to the protein-tyrosine phosphatase family. Non-receptor class 1 subfamily.
Contains 1 tyrosine-protein phosphatase domain. -
Post-translational
modificationsOxidized on Cys-215; the Cys-SOH formed in response to redox signaling reacts with the alpha-amido of the following residue to form a 4-amino-3-isothiazolidinone serine cross-link, triggering a conformational change that inhibits substrate binding and activity. The active site can be restored by reduction. -
Cellular localization
Endoplasmic reticulum membrane. - Information by UniProt
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Alternative names
- PTP1B
- Non receptor tyrosine phosphatase 1
- Protein phosphotyrosylphosphatase 1B
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260304 - Human PTPN1 knockout HeLa cell lysate 1 x 100µg ab255552 - 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 PTPN1 (PTP1B) knockout HeLa cell lysate (ab257617)Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: PTPN1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab252928 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab252928 Anti-PTP1B antibody [EPR22468-6] was shown to specifically react with PTP1B in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PTP1B knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab252928 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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. -
Western blot - Human PTPN1 (PTP1B) knockout HeLa cell lysate (ab257617)Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: PTPN1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab244207 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab244207 Anti-PTP1B antibody [EPR22474] was shown to specifically react with PTP1B in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab265014 (knockout cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PTP1B knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab244207 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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. -
Sanger Sequencing - Human PTPN1 knockout HeLa cell lysate (ab257617)Allele-1: 1 bp deletion in exon 4
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Sanger Sequencing - Human PTPN1 knockout HeLa cell lysate (ab257617)Allele-2: 1 bp insertion in exon 4
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Sanger Sequencing - Human PTPN1 knockout HeLa cell lysate (ab257617)Allele-3: Insertion of the selection cassette in exon 4
