IL1B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 47 bp deletion in exon 4.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 47 bp deletion in exon 4
Alternative names
Catabolin, H1, IFN beta inducing factor, IL 1, IL-1 beta, IL1B_HUMAN, IL1F2, Interleukin 1 beta precursor, Interleukin-1 beta, OAF, OTTHUMP00000162031, Osteoclast activating factor, Preinterleukin 1 beta, Preinterleukin beta, Pro interleukin 1 beta
Recommended products
IL1B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 47 bp deletion in exon 4.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 47 bp deletion in exon 4
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- IL1B
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- 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. 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.
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 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.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
5 product images
Immunocytochemistry/ Immunofluorescence - Human IL1B knockout THP-1 cell line (ab273762)
Anti-IL-1 beta antibody [EPR23851-127] ab254360 staining IL-1 beta in wild-type THP-1 cells (top panel) and IL1B knockout THP-1 cells (ab273762) (bottom panel) +/- TPA (80nM overnight) and LPS (100ng/ml, 6h). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-IL-1 beta antibody [EPR23851-127] ab254360 at 0.4μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Human IL1B knockout THP-1 cell line (ab273762)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-IL-1 beta antibody [EPR21086] ab216995 observed at 27-32 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.Anti-IL-1 beta antibody [EPR21086] ab216995 was shown to react with IL-1 beta in LPS-treated wild-type THP-1 cells in Western blot with loss of signal observed in IL1B knockout cell line ab273762 (LPS-treated) (knockout cell lysate Human IL1B knockout THP-1 cell lysate ab275508). Wild-type THP-1 and IL1B knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-IL-1 beta antibody [EPR21086] ab216995 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 (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 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-IL-1 beta antibody [EPR21086] (Anti-IL-1 beta antibody [EPR21086] ab216995) at 1/1000 dilution
Lane 1: Wild-type THP-1 untreated cell lysate at 30 µg
Lane 2: Wild-type THP-1 LPS-treated (3 h, 100 ng/ml) cell lysate at 30 µg
Lane 2: Western blot - Human IL1B knockout THP-1 cell line (ab273762)
Lane 3: IL1B knockout THP-1 untreated cell lysate at 30 µg
Lane 4: IL1B knockout THP-1 LPS-treated (3 h, 100 ng/ml) cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 27-32 kDa
Sandwich ELISA - Human IL1B knockout THP-1 cell line (ab273762)
Human IL-1 beta concentration was interpolated from the standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-1 beta ELISA kit (Human IL-1 beta ELISA Kit, Fluorescent ab229384). Wild-type and IL-1 beta knockout THP-1 cells (ab273762) were assessed in duplicate (n=2). Cells were either treated with LPS (100 ng/ml, 3 h) then ATP (5 mM, 45 min) to induce expression of IL-1 beta or not treated. Data are represented as the mean and error bars represent standard deviation.
Western blot - Human IL1B knockout THP-1 cell line (ab273762)
All lanes: Western blot - Anti-IL-1 beta antibody [RM1009] (Anti-IL-1 beta antibody [RM1009] ab283818) at 1/1000 dilution
Lane 1: Wild-type THP-1 Vehicle control: TPA (80 nM overnight) then LPS (0 ng/mL, 6 h) cell lysate at 20 µg
Lane 2: Wild-type THP-1 TPA (80 nM overnight) then LPS (100 ng/mL, 6 h) cell lysate at 20 µg
Lane 3: IL-1b knockout THP-1 Vehicle control: TPA (80 nM overnight) then LPS (0 ng/mL, 6 h) cell lysate at 20 µg
Lanes 3 - 4: Western blot - Human IL1B knockout THP-1 cell line (ab273762)
Lane 4: IL-1b knockout THP-1 TPA (80 nM overnight) then LPS (100 ng/mL, 6 h) cell lysate at 20 µg
Sanger Sequencing - Human IL1B knockout THP-1 cell line (ab273762)
Allele-1: 47 bp deletion in exon 4.
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