Human CCND1 (Cyclin D1) knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB261760)
Lanes 1- 2 : Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab16663 was shown to react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate ab256864) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16663 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 200 dilution and a 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-Cyclin D1 antibody [SP4] (<a href='/en-us/products/primary-antibodies/cyclin-d1-antibody-sp4-ab16663'>ab16663</a>) at 1/200 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CCND1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
Predicted band size: 34 kDa
Observed band size: 36 kDa
false
- WB
Lab
Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB261760)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : CCND1 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.
ab40754 Anti-Cyclin D1 antibody [EP272Y] was shown to specifically react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate ab256864) was used. Wild-type and Cyclin D1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40754 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CCND1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
Predicted band size: 34 kDa
Observed band size: 36 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB261760)
Homozygous : Insertion of the selection cassette in exon 1.
Complementary Products
Primary Antibodies
AB16663
BOND RX™ Validated|RabMAb|Recombinant|KO Validated|Lab Essentials
![Western blot - Anti-Cyclin D1 antibody [SP4] (AB16663)](https://content.abcam.com/products/images/cyclin-d1-antibody-sp4-ab16663--western-blot-img417814.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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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