Human DEGS1 (MLD) knockout HEK-293T cell lysate (ab257918)
Overview
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Product name
Human DEGS1 (MLD) knockout HEK-293T cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon1 and 1 bp insertion in exon1. -
Passage number
<20 -
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.
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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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262255 - Human DEGS1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Target
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Relevance
DEGS1 is a member of the membrane fatty acid desaturase family which is responsible for inserting double bonds into specific positions in fatty acids. It contains three His containing consensus motifs that are characteristic of a group of membrane fatty acid desaturases. It has sphingolipid-delta-4-desaturase activity and converts D-erythro-sphinganine to D-erythro-sphingosine (E-sphing-4-enine). -
Cellular localization
Endoplasmic reticulum and Mitochondrial -
Alternative names
- Degenerative spermatocyte homolog 1
- Degenerative spermatocyte homolog 1 lipid desaturase
- Degenerative spermatocyte homolog lipid desaturase
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab257918 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 38 kDa.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 38 kDa. |
Images
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Western blot - Human DEGS1 (MLD) knockout HEK293T cell lysate (ab257918)
Lane 1:Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2:DEGS1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3:U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate (20 ug)
Lane 4:T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate (20 ug)ab167169 was shown to specifically react with MLD in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266481 (knockout cell lysate ab257918) was used. Wild-type and MLD knockout samples were subjected to SDS-PAGE. ab167169 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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Western blot - Human DEGS1 (MLD) knockout HEK293T cell lysate (ab257918)Lane 1:Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2:DEGS1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3:U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate (20 ug)
Lane 4:T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate (20 ug)ab185237 was shown to specifically react with MLD in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266481 (knockout cell lysate ab257918) was used. Wild-type and MLD knockout samples were subjected to SDS-PAGE. ab185237 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 DEGS1 knockout HEK293T cell lysate (ab257918)Allele-1: 14 bp deletion in exon1
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Sanger Sequencing - Human DEGS1 knockout HEK293T cell lysate (ab257918)Allele-2: 1 bp insertion in exon1
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab257918 has not yet been referenced specifically in any publications.