Human RTN3 (RTN3/HAP) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)
False colour image of Western blot : Anti-RTN3/HAP antibody staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab68328 was shown to bind specifically to RTN3/HAP. A band was observed at 24 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in RTN3 knockout cell line ab266517 (knockout cell lysate ab258647). To generate this image, wild-type and RTN3 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-RTN3/HAP antibody (<a href='/en-us/products/primary-antibodies/rtn3-hap-antibody-ab68328'>ab68328</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human RTN3 (RTN3/HAP) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-rtn3-rtn3-hap-knockout-hek-293t-cell-lysate-ab258647'>ab258647</a>) at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
U-87 MG cell lysate at 20 µg
Predicted band size: 113 kDa
Observed band size: 24 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RTN3 (RTN3/HAP) knockout HEK-293T cell line (AB266517)
Homozygous : 1 bp insertion in exon 1
Complementary Products
Primary Antibodies
AB250023
Anti-Nogo antibody [EPR12266] - BSA and Azide free
RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-Nogo antibody [EPR12266] - BSA and Azide free (AB250023)](https://content.abcam.com/products/images/nogo-antibody-epr12266-bsa-and-azide-free-ab250023--flow-cytometry-intracellular-img26400.jpg)
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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
Zygosity
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
The RTN3/HAP protein can influence neurite outgrowth implicating it in the development and maintenance of the nervous system. This protein associates closely with other members of the reticulon family forming complexes important for its functional roles within the cell. RTN3/HAP has been implicated in mechanisms that protect neurons from stress especially related to endoplasmic reticulum stress by its participation in membrane remodeling and regulation of lipid metabolism.
Pathways
Data shows that RTN3/HAP interacts with components of the endoplasmic reticulum stress response pathway and lipid biosynthesis pathways. It works alongside proteins such as RTN4/Nogo contributing to the regulation of axon guidance and regeneration. These pathways are essential for maintaining cellular homeostasis and responding to physiological stress with RTN3/HAP helping adapt or protect cells under challenging conditions.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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