CHMP2B KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%
Alternative names
ALS17, CHM2B_HUMAN, CHMP family, member 2B, CHMP2.5, Charged multivesicular body protein 2b, Chromatin-modifying protein 2b, VPS2 homolog B, VPS2B, Vacuolar protein sorting 2, yeast, homolog of, B, Vacuolar protein sorting 2-2, Vacuolar protein sorting-associated protein 2-2, Vps2-2, hVps2-2
Recommended products
CHMP2B KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- CHMP2B
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
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Full details and terms and conditions can be found here:
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3 product images
Western blot - Human CHMP2B knockout A549 cell line (ab261874)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CHMP2B antibody ab33174 observed at 30 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-CHMP2B antibody ab33174 was shown to recognize CHMP2B in wild-type A549 cells as signal was lost at the expected MW in CHMP2B knockout cell line ab261874 (knockout cell lysate Human CHMP2B knockout A549 cell lysate ab261683). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and CHMP2B knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-CHMP2B antibody ab33174 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CHMP2B antibody (Anti-CHMP2B antibody ab33174) at 1 µg/mL
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: CHMP2B knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human CHMP2B knockout A549 cell line (ab261874)
Lane 3: A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 24 kDa
Western blot - Human CHMP2B knockout A549 cell line (ab261874)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CHMP2B antibody [EPR10807(B)] ab157208 observed at 45 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
Anti-CHMP2B antibody [EPR10807(B)] ab157208 was shown to specifically react with CHMP2B in wild-type A549 cells as signal was lost in CHMP2B knockout cell line ab261874 (knockout cell lysate Human CHMP2B knockout A549 cell lysate ab261683). Wild-type and CHMP2B knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-CHMP2B antibody [EPR10807(B)] ab157208 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CHMP2B antibody [EPR10807(B)] (Anti-CHMP2B antibody [EPR10807(B)] ab157208) at 1/1000 dilution
Lane 1: Wild-type A549 whole cell lysate at 20 µg
Lane 2: CHMP2B knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human CHMP2B knockout A549 cell line (ab261874)
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 24 kDa
Next Generation Sequencing - Human CHMP2B knockout A549 cell line (ab261874)
X = 1 bp insertion
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