JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB266481

Human DEGS1 (MLD) knockout HEK-293T cell line

Be the first to review this product! Submit a review

|

(1 Publication)

DEGS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)
  • WB

Lab

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)

Lanes 1-4 : Merged signal (red and green). Green - ab185237 observed at 38 kDa. Red - loading control ab8245 observed at 36 kDa.

ab185237 Anti-MLD antibody [EPR9680] 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 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MLD antibody [EPR9680] (<a href='/en-us/products/primary-antibodies/mld-antibody-epr9680-ab185237'>ab185237</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

DEGS1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (ab266481)

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

T-47D (Human ductal breast epithelial tumor cell line) whole 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: 38 kDa

Observed band size: 38 kDa

false

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)
  • WB

Lab

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)

Lanes 1-4 : Merged signal (red and green). Green - ab167169 observed at 38 kDa. Red - loading control ab8245 observed at 36 kDa.

ab167169 Anti-MLD antibody [EPR9681] 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 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MLD antibody [EPR9681] (<a href='/en-us/products/primary-antibodies/mld-antibody-epr9681-ab167169'>ab167169</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

DEGS1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human DEGS1 (MLD) knockout HEK-293T cell line (ab266481)

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

T-47D (Human ductal breast epithelial tumor cell line) whole 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: 38 kDa

Observed band size: 38 kDa

false

Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)
  • Sanger seq

Unknown

Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)

Allele-1 : 14 bp deletion in exon1

Cell Culture - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)
  • Cell Culture

Lab

Cell Culture - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)

Representative images DEGS1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)
  • Sanger seq

Unknown

Sanger Sequencing - Human DEGS1 (MLD) knockout HEK-293T cell line (AB266481)

Allele-2 : 1 bp insertion in exon 1.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 1 and 1 bp insertion in exon 1

style
left-column

Complementary Products

Primary Antibodies

AB167169

Anti-MLD antibody [EPR9681]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLD antibody [EPR9681] (AB167169)

  • 1
  • 2
Proteins & Peptides

AB160100

Recombinant Human MLD protein

SDS-PAGE - Recombinant Human MLD protein (AB160100)

  • 0
  • 0
Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

Mass Spectrometry - Recombinant Human WFDC2 Protein (His tag) (AB313361)

  • 0
  • 0
Proteins & Peptides

AB195163

Recombinant Human ZMYND8 protein

SDS-PAGE - Recombinant Human ZMYND8 protein (AB195163)

  • 0
  • 0
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

We will provide viable cells that proliferate on revival.

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.

style
left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266481-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266481-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab266481 Human DEGS1 (MLD) knockout HEK-293T cell line", "number":"AB266481-CMP01" } ] }, "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266481-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266481 Human DEGS1 (MLD) knockout HEK-293T cell line", "number":"AB266481-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266481-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266481-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266481 Human DEGS1 (MLD) knockout HEK-293T cell line", "number":"AB266481-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266481-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266481 Human DEGS1 (MLD) knockout HEK-293T cell line", "number":"AB266481-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
DEGS1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
style
left-column

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MLD also known as Metachromatic Leukodystrophy protein is an enzyme of significant importance in cellular function. This protein has a molecular mass of around 50 kDa. It is primarily expressed in the brain but is also found in peripheral tissues. MLD breaks down sulfatides which are important components of myelin sheaths. The myelin sheaths encapsulate nerve cells and are essential for efficient nerve signal transmission.
Biological function summary

Metachromatic Leukodystrophy protein plays an important role in the maintenance of the nervous system structure and function. It is involved in the degradation of certain lipid compounds to prevent toxic accumulation. When the function of the MLD protein is impaired sulfatides accumulate leading to cellular damage and functional deficits. MLD does not generally function as part of a larger complex; instead it acts independently within its cellular environment.

Pathways

Metachromatic Leukodystrophy protein is integral to the sphingolipid degradation pathway. This pathway includes the catabolism of complex lipid molecules preventing harmful buildup within cells. Another enzyme arylsulfatase A closely interacts with the MLD protein and participates in the same pathway further demonstrating its importance in managing cellular lipids and maintaining cell health.

Metachromatic Leukodystrophy protein is directly related to Metachromatic Leukodystrophy a genetic disorder affecting the nervous system. This disease results from mutations in the gene that encodes for the MLD protein leading to demyelination and neurological decline. Other proteins like cerebroside sulfate become dysregulated in this disorder indicating a complex network of biomolecular interactions disrupted when MLD function is compromised.
style
left-column
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column

Target data

See full target information DEGS1
style
left-column

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Journal of virology 95:e0080721 PubMed34106748

2021

-(4-Hydroxyphenyl) Retinamide Suppresses SARS-CoV-2 Spike Protein-Mediated Cell-Cell Fusion by a Dihydroceramide Δ4-Desaturase 1-Independent Mechanism.

Applications

Unspecified application

Species

Unspecified reactive species

Yasuhiro Hayashi,Kiyoto Tsuchiya,Mizuki Yamamoto,Yoko Nemoto-Sasaki,Kazunari Tanigawa,Kotaro Hama,Yusuke Ueda,Takashi Tanikawa,Jin Gohda,Kenji Maeda,Jun-Ichiro Inoue,Atsushi Yamashita
View all publications
style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com