Human SAMD9 knockout A549 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human SAMD9 knockout A549 cell line (AB267038)
Lanes 1-4 : Merged signal (red and green). Green - ab180575 observed at 184 kDa. Red - loading control ab8245 observed at 36 kDa.
ab180575 Anti-SAMD9 antibody [EPR13603] was shown to specifically react with SAMD9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267038 (knockout cell lysate ab257656) was used. Wild-type and SAMD9 knockout samples were subjected to SDS-PAGE. ab180575 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-SAMD9 antibody [EPR13603] (<a href='/en-us/products/primary-antibodies/samd9-antibody-epr13603-ab180575'>ab180575</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
SAMD9 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human SAMD9 knockout A549 cell line (ab267038)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
Human skeletal muscle 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: 184 kDa
Observed band size: 184 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SAMD9 knockout A549 cell line (AB267038)
Homozygous : 1 bp insertion in exon2
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Cell Lines & Lysates
AB266215
Human NME1 (NM23A) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
SAMD9 takes part in controlling cellular proliferation and inhibiting abnormal growth mostly acting as a growth suppressor. Its functions link to maintaining homeostasis within cells and aiding in preventing uncontrolled cell expansion. SAMD9 functions independently of any known stable protein complex performing its regulatory role in diverse cellular contexts. The protein's involvement spans multiple cell types reflecting its broad impact on normal human physiology.
Pathways
SAMD9 interacts with critical signaling modules such as the mTOR pathway and the interferon signaling pathway. It modulates cellular responses related to growth control and stress adaptation. Interactions with other proteins like TSC1 and TSC2 within the mTOR pathway highlight SAMD9's engagement in regulating cellular responses to environmental changes and nutrient availability.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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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