Human KDM1B (LSD2 / AOF1) knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
KDM1B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 3 and 5 bp deletion in exon 3.
View Alternative Names
AOF1, C6orf193, FLJ33898, FLJ34109, FLJ43328, Flavin-containing amine oxidase domain-containing protein 1, KDM1B_HUMAN, LSD2, Lysine-specific demethylase 2, Lysine-specific histone demethylase 1B, Lysine-specific histone demethylase 2, amine oxidase (flavin containing) domain 1, amine oxidase, flavin containing 1, bA204B7.3, dJ298J15.2, lysine (K)-specific demethylase 1B
- WB
Lab
Western blot - Human KDM1B (LSD2 / AOF1) knockout HeLa cell line (AB265969)
Lanes 1-4 : Merged signal (red and green). Green - ab193080 observed at 95 kDa. Red - loading control ab8245 observed at 36 kDa.
ab193080 Anti-LSD2 / AOF1 antibody [EPR18508] was shown to specifically react with LSD2 / AOF1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265969 (knockout cell lysate ab258016) was used. Wild-type and LSD2 / AOF1 knockout samples were subjected to SDS-PAGE. ab193080 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-LSD2 / AOF1 antibody [EPR18508] (<a href='/en-us/products/primary-antibodies/lsd2-aof1-antibody-epr18508-ab193080'>ab193080</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
KDM1B knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human KDM1B (LSD2 / AOF1) knockout HeLa cell line (ab265969)
Lane 3:
HAP1 cell lysate at 20 µg
Lane 4:
HCT116 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: 92 kDa
Observed band size: 95 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human KDM1B (LSD2 / AOF1) knockout HeLa cell line (AB265969)
Allele-1 : 5 bp deletion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human KDM1B (LSD2 / AOF1) knockout HeLa cell line (AB265969)
Allele-2 : 2 bp deletion in exon 3.
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 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 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
LSD2 impacts gene expression by altering the epigenetic state of chromatin. It is not part of a well-defined large multiprotein complex allowing LSD2 to directly interact with histones. Through its demethylase activity it modulates the transcriptional regulation impacting cellular differentiation and proliferation. This makes it an important factor in maintaining cellular homeostasis.
Pathways
LSD2 is involved in histone modification processes particularly through the demethylation of H3K4me1 and H3K4me2 marks. These changes are important in the regulation of gene expression patterns. LSD2 often interacts with the REST corepressor complex affecting the expression of genes involved in neural development and function. It plays a role in pathways related to cellular stress responses and metabolic regulation highlighting its importance in cellular adaptation mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB193080
Anti-LSD2 / AOF1 antibody [EPR18508]
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-LSD2 / AOF1 antibody [EPR18508] (AB193080)](https://content.abcam.com/products/images/lsd2-aof1-antibody-epr18508-ab193080--immunocytochemistry-immunofluorescence-img68272.jpg)
- 0
- 0
Cell Lines & Lysates
AB266158
Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
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