Human SH3GL1 (EEN) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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SH3GL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 2 and Insertion of the selection cassette in exon 2. 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
CNSA1, EEN, EEN fusion partner of MLL, Endophilin-2, Endophilin-A2, Extra 11 19 leukemia fusion gene, Extra eleven-nineteen leukemia fusion gene protein, Fusion partner of MLL, MGC111371, SH3 containing Grb 2 like 1 protein, SH3 containing protein EEN, SH3 domain GRB2 like 1, SH3 domain protein 2B, SH3 domain-containing GRB2-like protein 1, SH3D2B, SH3G1_HUMAN, SH3P8, Sh3gl1
- WB
Lab
Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell line (AB266260)
Lanes 1 - 4 : Merged signal (red and green). Green - Mouse monoclonal antibody observed at 48 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
Mouse monoclonal antibody was shown to react with Endophilin II in wild-type HEK-293T cells in Western blot with loss of signal observed in SH3GL1 knockout cell line ab266260 (SH3GL1 knockout cell lysate ab259109). Wild-type HEK-293T and SH3GL1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Mouse monoclonal antibody and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 100 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:
Mouse monoclonal antibody at 1/100 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
SH3GL1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell line (ab266260)
Lane 3:
HUVEC cell lysate at 20 µg
Lane 4:
A-431 cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell line (AB266260)
Allele-1 : 10 bp deletion in exon 2
- Sanger seq
Unknown
Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell line (AB266260)
Allele-2 : Insertion of the selection cassette in exon 2.
Reactivity data
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.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
EEN participates in the formation of protein complexes necessary for synaptic vesicle recycling important for neurotransmitter release in neurons. It binds to proline-rich motifs on its partner proteins which helps in vesicle scission during endocytosis. These interactions support vesicle fission and transport making EEN important for maintaining efficient synaptic transmission. Apart from neurons EEN also contributes to endocytic processes in other cell types indicating its wider functional role.
Pathways
EEN plays a significant role in the regulation of intracellular trafficking pathways and is a part of the endocytic pathway. It links with proteins such as dynamin and clathrin integral components of the endocytic machinery that facilitate vesicle budding and release. These interactions ensure proper recycling of synaptic vesicles and membrane components vital for cellular homeostasis. Collaborations with proteins like Bin1 show EEN's involvement in coordinating endocytosis and membrane curvature recognition.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Cell Lines & Lysates
AB266287
Human RRAGD (Rag D) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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