Human GLS knockout MCF7 cell line
Be the first to review this product! Submit a review
|
(0 Publication)
GLS KO cell line available to order. KO validated. Free of charge wild type control available. 137 bp deletion after Leu31. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
K-glutaminase, GLS1, L-glutamine amidohydrolase, GLS, KIAA0838
- NGS
Lab
Next Generation Sequencing - Human GLS knockout MCF7 cell line (AB287444)
137 bp deletion after Leu31
- WB
Lab
Western blot - Human GLS knockout MCF7 cell line (AB287444)
Western blot : Anti-Glutaminase antibody [EP7212] ab156876 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. Three specific bands are detected by ab156876. We believe these correspond to isoform 1 (KGA), isoform 3 (GAC) and the mature mitochondrial subunits. Bands were observed at 68, 60 and 55 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in GLS knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-Glutaminase antibody [EP7212] (<a href='/en-us/products/primary-antibodies/glutaminase-antibody-ep7212-ab156876'>ab156876</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysates at 20 µg
Lane 2:
Western blot - Human GLS knockout MCF7 cell line (ab287444) at 20 µg
Lane 3:
HeLa cell lysates at 20 µg
Lane 4:
OVCAR-3 cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 68 kDa,60 kDa,55 kDa
false
- WB
Lab
Western blot - Human GLS knockout MCF7 cell line (AB287444)
Western blot : Anti-Glutaminase C antibody [EPR19525] ab202027 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in GLS knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-Glutaminase C antibody [EPR19525] (<a href='/en-us/products/primary-antibodies/glutaminase-c-antibody-epr19525-ab202027'>ab202027</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysates at 20 µg
Lane 2:
Western blot - Human GLS knockout MCF7 cell line (ab287444) at 20 µg
Lane 3:
HeLa cell lysates at 20 µg
Lane 4:
OVCAR-3 cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 60 kDa
false
Complementary Products
Proteins & Peptides
AB126670
Recombinant Human Junctional Adhesion Molecule 1/JAM-A protein (His tag N-Terminus)
- 0
- 0
Primary Antibodies
AB124930
Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)]
RabMAb|Recombinant|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)](https://content.abcam.com/products/images/igf2bp2-imp-2-antibody-epr6741b-ab124930--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img139417.jpg)
- 4
- 1
Primary Antibodies
AB154785
Anti-Rho guanine exchange factor 16/NBR antibody [EPR9950]
RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho guanine exchange factor 16/NBR antibody [EPR9950] (AB154785)](https://content.abcam.com/products/images/rho-guanine-exchange-factor-16-nbr-antibody-epr9950-ab154785--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img40687.jpg)
- 0
- 0
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type MCF7 cell line (ab288560). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Glutaminase plays a central role in cellular processes by facilitating the production of glutamate which is an important neurotransmitter in the brain and a precursor for the synthesis of gamma-aminobutyric acid (GABA). It is not part of a larger complex but acts independently in the mitochondrial matrix where it performs the glutaminase reaction. The activity of this enzyme is monitored closely because it affects multiple cellular functions and has implications in many diseases.
Pathways
Glutaminase is instrumental in the glutaminolysis and tricarboxylic acid (TCA) cycles. These pathways are essential for cellular energy production and biosynthesis. GLS1 activity influences other proteins such as alanine transaminase (ALT) and aspartate transaminase (AST) which also play roles in amino acid metabolism and energy production. Glutaminase inhibitors have been explored as potential therapeutic agents by disrupting the glutaminolysis pathway in cancer cells leading to decreased tumor growth.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
Subculture guidelines
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Target data
Product promise
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