Human Lgals9 (galectin 9/Gal-9) knockout THP-1 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human Lgals9 (galectin 9/Gal-9) knockout THP-1 cell line (AB269505)
Lanes 1 - 2 : Merged signal (red and green). Green - ab227046 observed at 35 kDa. Red - loading control Mouse anti Histone H3 observed at 18 kDa.
ab227046 was shown to react with galectin 9/Gal-9 in wild-type THP-1 cells in Western blot with loss of signal observed in LGALS9 knockout cell line ab269505 (knockout cell lysate ab269667). Wild-type THP-1 and LGALS9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab227046 and Mouse anti Histone H3 overnight at 4 ° 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 h at room temperature before imaging.
All lanes:
Western blot - Anti-galectin 9/Gal-9 antibody [EPR22214] (<a href='/en-us/products/primary-antibodies/galectin-9-gal-9-antibody-epr22214-ab227046'>ab227046</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
LGALS9 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human Lgals9 (galectin 9/Gal-9) knockout THP-1 cell line (ab269505)
Predicted band size: 40 kDa
Observed band size: 35 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human Lgals9 (galectin 9/Gal-9) knockout THP-1 cell line (AB269505)
2 bp deletion after Gln25 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human Lgals9 (galectin 9/Gal-9) knockout THP-1 cell line (AB269505)
Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift : 98%
Complementary Products
Primary Antibodies
AB227046
Anti-galectin 9/Gal-9 antibody [EPR22214]
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-galectin 9/Gal-9 antibody [EPR22214] (AB227046)](https://content.abcam.com/products/images/galectin-9-gal-9-antibody-epr22214-ab227046--immunocytochemistry-immunofluorescence-img76996.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 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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
Galectins like galectin-9 play essential roles in regulating diverse immune responses. Galectin-9 acts within the immune system by modulating inflammation and apoptosis. It acts as both a pro-apoptotic factor for T cells and a protein mediator in innate immune functions. Galectin-9 proteins can form complexes with other lectins and cell surface receptors contributing to cellular signaling processes.
Pathways
Galectin-9 is integral to the immune system's functional networks. It participates in the T-cell regulation pathways and the innate immunity pathways. In these pathways it interacts with other proteins like TIM-3 which mediates T-cell apoptosis and contributes to immune tolerance. These interaction networks play important roles in maintaining the body's immune homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
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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