Human ALDH2 knockout SW480 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human ALDH2 knockout SW480 cell line (AB270483)
Lanes 1 - 4 : Merged signal (red and green). Green - ab133306 observed at 53 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab133306 was shown to react with ALDH2 in wild-type SW480 cells in western blot Loss of signal was observed when ALDH2 knockout cell line ab270483 (knockout cell lysate ab270506) was used. Wild-type and ALDH2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab133306 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ALDH2 antibody [EPR4494] (<a href='/en-us/products/primary-antibodies/aldh2-antibody-epr4494-ab133306'>ab133306</a>) at 1/1000 dilution
Lane 1:
Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 2:
ALDH2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human ALDH2 knockout SW480 cell line (ab270483)
Lane 3:
Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 56 kDa
Observed band size: 53 kDa,56 kDa
false
- NGS
Lab
Next Generation Sequencing - Human ALDH2 knockout SW480 cell line (AB270483)
2 bp deletion after Leu107 of the WT protein
Complementary Products
![Western blot - Anti-ALDH2 antibody [EPR4494] (AB133306)](https://content.abcam.com/products/images/aldh2-antibody-epr4494-ab133306--western-blot-img2269.jpg)
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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 2-3x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Slow growing. A partial media change is recommended at least twice between passages.
Culture medium
Ham's F-12 + 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
ALDH2 serves a detoxifying function by preventing oxidative stress and promoting cellular health. As part of the larger aldehyde dehydrogenase family this enzyme forms homotetramer complexes to efficiently catalyze the oxidation of aldehydes. ALDH2 supplementation may enhance its activity in individuals with reduced function due to genetic polymorphisms providing potential therapeutic benefits.
Pathways
ALDH2 integrates into the alcohol metabolism and retinoic acid signaling pathways. In alcohol metabolism it collaborates with alcohol dehydrogenase enzymes such as ADH1 and ADH2 to process alcohol consumption compounds. In retinoic acid signaling ALDH2 helps regulate the biosynthesis of retinoic acid influencing cell proliferation and differentiation processes.
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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