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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.

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Images

Immunocytochemistry/ Immunofluorescence - Human IL1B knockout THP-1 cell line (AB273762), expandable thumbnail
  • Western blot - Human IL1B knockout THP-1 cell line (AB273762), expandable thumbnail
  • Sandwich ELISA - Human IL1B knockout THP-1 cell line (AB273762), expandable thumbnail
  • Western blot - Human IL1B knockout THP-1 cell line (AB273762), expandable thumbnail
  • Sanger Sequencing - Human IL1B knockout THP-1 cell line (AB273762), expandable thumbnail

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

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
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 47 bp deletion in exon 4
Disease
Acute Monocytic Leukemia
Concentration
Loading...

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

Recommended control: Human wild-type THP-1 cell line (ab275477). 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.
  • Upon thawing make banks as soon as possible and use each bank within 10-20 passages
  • As cell line growth can vary please attempt culture for 2 weeks from revival of initial bank before contacting the technical team.
  • 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.

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    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:
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    5 product images

    • Immunocytochemistry/ Immunofluorescence - Human IL1B knockout THP-1 cell line (ab273762), expandable thumbnail

      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), expandable thumbnail

      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), expandable thumbnail

      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), expandable thumbnail

      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), expandable thumbnail

      Sanger Sequencing - Human IL1B knockout THP-1 cell line (ab273762)

      Allele-1: 47 bp deletion in exon 4.

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