Human SNAP29 knockout HeLa cell line
- Advanced Validation
- What is this?
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SNAP29 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 5 bp deletion in exon 1.
View Alternative Names
CEDNIK, FLJ21051, SNP29_HUMAN, Soluble 29 kDa NSF attachment protein, Synaptosomal associated protein 29kDa, Synaptosomal-associated protein 29, Vesicle-membrane fusion protein SNAP-29
- WB
Lab
Western blot - Human SNAP29 knockout HeLa cell line (AB265289)
Lanes 1-3 : Merged signal (red and green). Green - ab181151 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.
ab181151 Anti-SNAP29 antibody [EPR9198(2)] was shown to specifically react with SNAP29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265289 (knockout cell lysate ab257693) was used. Wild-type and SNAP29 knockout samples were subjected to SDS-PAGE. ab181151 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-SNAP29 antibody [EPR9198(2)] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr91982-ab181151'>ab181151</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SNAP29 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SNAP29 knockout HeLa cell line (ab265289)
Lane 3:
HepG2 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: 29 kDa
Observed band size: 29 kDa
false
- WB
Lab
Western blot - Human SNAP29 knockout HeLa cell line (AB265289)
Lanes 1-3 : Merged signal (red and green). Green - ab138500 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.
ab138500 Anti-SNAP29 antibody [EPR9199] was shown to specifically react with SNAP29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265289 (knockout cell lysate ab257693) was used. Wild-type and SNAP29 knockout samples were subjected to SDS-PAGE. ab138500 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-SNAP29 antibody [EPR9199] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr9199-ab138500'>ab138500</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SNAP29 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SNAP29 knockout HeLa cell line (ab265289)
Lane 3:
HepG2 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: 29 kDa
Observed band size: 29 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SNAP29 knockout HeLa cell line (AB265289)
Allele-2 : 1 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human SNAP29 knockout HeLa cell line (AB265289)
Allele-1 : 5 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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
RNA encoding this protein facilitates vesicle-mediated transport on the cellular level especially under conditions requiring rapid membrane addition or reshaping. SNAP29 acts alongside other SNARE proteins like syntaxin and VAMP in the SNARE complex to modulate membrane interactions. The protein eases the process of cytoplasmic content exchange between organelles. As part of these complexes SNAP29 maintains cellular homeostasis by contributing to membrane dynamics and trafficking processes.
Pathways
Vesicular transport in the cell highly depends on SNAP29 especially in exocytosis and endocytosis pathways. Alongside other SNARE proteins SNAP29 fits into the vesicle docking and fusion pathways ensuring efficient membrane recycling and cargo delivery. It interacts with several proteins in the process including syntaxin 1 and VAMP2 to facilitate these important functions within these pathways.
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
![Western blot - Anti-SNAP29 antibody [EPR9198(2)] (AB181151)](https://content.abcam.com/products/images/snap29-antibody-epr91982-ab181151--western-blot-img612.jpg)
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