Human GPI (AMF) knockout HEK-293T cell lysate
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GPI KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 76 bp deletion in exon2.
View Alternative Names
AMF, Aurocrine motility factor, Autocrine motility factor, DKFZp686C13233, EC 5.3.1.9, G6PI_HUMAN, GNPI, GPI, Glucose phosphate isomerase, Glucose-6-phosphate isomerase, Gpi1, Hexose monophosphate isomerase, Hexosephosphate isomerase, Neuroleukin, Oxoisomerase, PGI, PHI, Phosphoglucose isomerase, Phosphohexomutase, Phosphohexose isomerase, Phosphosaccharomutase, SA-36, Sperm antigen 36
- WB
Lab
Western blot - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)
Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : GPI knockout HEK293T cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)
ab167394 was shown to specifically react with AMF in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266834 (knockout cell lysate ab257458) was used. Wild-type and AMF knockout samples were subjected to SDS-PAGE. ab167394 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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.
All lanes:
Western blot - Anti-AMF antibody [EPR11663(B)] (<a href='/en-us/products/primary-antibodies/amf-antibody-epr11663b-ab167394'>ab167394</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
GPI knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human GPI (AMF) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gpi-amf-knockout-hek-293t-cell-line-ab266834'>ab266834</a>)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 63 kDa
Observed band size: 63 kDa
false
- WB
Lab
Western blot - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)
Lane 1 : wild-type HEK-293T cell lysate 20 ug
Lane 2 : GPI knockout HEK-293T cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab66340 observed at 63 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab66340 was shown to react with AMF in wild-type HEK-293T cells in Western blot with loss of signal observed in GPI knockout cell line ab266834 (GPI knockout cell lysate ab257458). Wild-type HEK-293T and GPI knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab66340 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-AMF antibody [1B7D7] (<a href='/en-us/products/primary-antibodies/amf-antibody-1b7d7-ab66340'>ab66340</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
GPI knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human GPI (AMF) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gpi-amf-knockout-hek-293t-cell-line-ab266834'>ab266834</a>)
Predicted band size: 63 kDa
Observed band size: 63 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)
Homozygous : 76 bp deletion in exon2
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
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.
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. 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.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Primary Antibodies
AB313590
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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