Human HOXA9 knockout HCT116 cell line
- Advanced Validation
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HOXA9 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
ABD-B, Abd-B, drosophila, homolog of, D6a9, HOX1, HOXA9/MSI2 fusion gene, included, HOXA9/NUP98 fusion gene, included, HXA9_HUMAN, Homeobox 1G, Homeobox A9, Homeobox protein Hox-1.7, Homeobox protein Hox-1G, Homeobox protein Hox-A9, Homeodomain protein HOXA9, Hox-1.7, Hox-1.7, mouse, homolog of, Hox1G, Hox1r5, MGC1934
- WB
Lab
Western blot - Human HOXA9 knockout HCT116 cell line (AB287488)
Western blot : Anti-HOXA9 antibody [EPR3655(2)] (ab140631) staining at 1/1000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab140631 was shown to bind specifically to HOXA9. A band was observed at 30 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in HOXA9 knockout cell line. To generate this image, wild-type and HOXA9 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 1 minute exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-HOXA9 antibody [EPR3655(2)] (<a href='/en-us/products/primary-antibodies/hoxa9-antibody-epr36552-ab140631'>ab140631</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human HOXA9 knockout HCT116 cell line (ab287488)
Lane 2:
HOXA9 knockout HCT 116 cell lysate at 20 µg
Secondary
Lanes 1 - 2:
HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution
Lanes 1 - 2:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 30 kDa
false
- NGS
Lab
Next Generation Sequencing - Human HOXA9 knockout HCT116 cell line (AB287488)
191 bp deletion (allele 1) and 179 bp deletion (allele 2) in exon 1, CCDS5409.1
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab288559). 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.
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
McCoY5a + 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
HOXA9 controls the transition and lineage commitment of hematopoietic stem cells. The protein often interacts as part of a transcriptional complex involving other HOX family proteins to maintain proper gene expression balance. This regulatory function is essential for normal gene expression patterns needed in stem cell differentiation and organ development processes.
Pathways
HOXA9 plays significant roles in the hematopoietic cell lineage and the retinoic acid signaling pathway. Through these pathways it collaborates with other HOX proteins like HOXB4 to maintain stem cell properties and influence the differentiation process. These interactions facilitate the production of necessary blood cell types and ensure effective response mechanisms to various developmental cues.
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
Complementary Products
![Immunoprecipitation - Anti-HOXA9 antibody [EPR3655(2)] (AB140631)](https://content.abcam.com/products/images/hoxa9-antibody-epr36552-ab140631--immunoprecipitation-img408403.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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