Human SORT1 (Sortilin/NT3) knockout HeLa cell line
- Advanced Validation
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SORT1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 5 and 16 bp deletion in exon 5 and 1 bp insertion in exon 5 and Insertion of the selection cassette in exon 5.
View Alternative Names
100 kDa NT receptor, Glycoprotein 95, Gp 95, LDLCQ6, NT-3, NTR 3, Neurotensin receptor 3, OTTHUMP00000013784, SORT 1, SORT1 (gene name), SORT_HUMAN, Sortilin, Sortilin 1
- WB
Lab
Western blot - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Lane 1 : Wild-type HeLa cell lysate (20µg)
Lane 2 : SORT1 knockout HeLa cell lysate (20µg)
Lanes 1- 2 : Merged signal (red and green). Green - ab188586 observed at 100 kDa. Red - loading control ab8245 observed at 37 kDa.
ab188586 Anti-Sortilin/NT3 antibody [EPR15010] was shown to specifically react with Sortilin/NT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264772 (knockout cell lysate ab257696) was used. Wild-type and Sortilin/NT3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab188586 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-Sortilin/NT3 antibody [EPR15010] (<a href='/en-us/products/primary-antibodies/sortilin-nt3-antibody-epr15010-ab188586'>ab188586</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SORT1 (Sortilin/NT3) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-sort1-sortilin-nt3-knockout-hela-cell-lysate-ab257696'>ab257696</a>) at 20 µg
Secondary
Lanes 1 - 2:
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 1/20000 dilution
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 92 kDa
Observed band size: 100 kDa
false
- WB
Unknown
Western blot - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Lanes 1- 2 : Merged signal (red and green). Green - ab16640 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab16640 was shown to react with Sortilin/NT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264772 (knockout cell lysate ab257696) was used. Wild-type HeLa and SORT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16640 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 μg/ml and a 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-Sortilin/NT3 antibody (<a href='/en-us/products/primary-antibodies/sortilin-nt3-antibody-ab16640'>ab16640</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SORT1 (Sortilin/NT3) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-sort1-sortilin-nt3-knockout-hela-cell-lysate-ab257696'>ab257696</a>) at 20 µg
Secondary
Lanes 1 - 2:
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 1/20000 dilution
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 92 kDa
Observed band size: 100 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Allele-2 : 11 bp deletion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Allele-4 : Insertion of the selection cassette in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Allele-1 : 16 bp deletion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human SORT1 (Sortilin/NT3) knockout HeLa cell line (AB264772)
Allele-3 : 1 bp insertion in exon 5.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). 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, 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
NT-3 protein influences the development and differentiation of the nervous system. Sortilin serves as a receptor for NT-3 facilitating the interaction with the TrkC receptor on neuronal cells. Sortilin forms a complex with NT-3 contributing to critical cellular processes. Being part of this receptor complex Sortilin modulates neuron-related signaling pathways which are essential for neurogenesis and synaptic plasticity.
Pathways
Sortilin and NT-3 play significant roles in the neurotrophin signaling pathway and influence the MAPK signaling cascade. These pathways are pivotal in neuron survival differentiation and growth. NT-3 interacts with other proteins such as TrkC and p75 to propagate these signals. Through its association with TrkC receptor Sortilin mediates critical cellular responses linked to neurotrophin signaling. The interplay between these proteins allows precise regulation of neuronal activities in various physiological states.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
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