Human CASP1 (Caspase-1) knockout THP-1 cell line (ab276116)
Overview
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Product name
Human CASP1 (Caspase-1) knockout THP-1 cell line -
Parental Cell Line
THP-1 -
Organism
Human -
Passage number
<20 -
Knockout validation
Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
1 -
General notes
Recommended control: Human wild-type THP-1 cell line (ab281894). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: RPMI + 10% FBS + 0.05 mM β-mercaptoethanol
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 2-4x104 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.Subculture guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2-4x105 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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
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Number of cells
1 x 106 cells/vial, 1 mL -
Viability
~80% -
Adherent /Suspension
Suspension -
Tissue
Blood -
Cell type
acute monocytic leukemia -
Disease
Acute Monocytic Leukemia -
Gender
Male -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
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Function
Thiol protease that cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes. Important for defense against pathogens. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Can also promote apoptosis. -
Tissue specificity
Expressed in larger amounts in spleen and lung. Detected in liver, heart, small intestine, colon, thymus, prostate, skeletal muscle, peripheral blood leukocytes, kidney and testis. No expression in the brain. -
Sequence similarities
Belongs to the peptidase C14A family.
Contains 1 CARD domain. -
Post-translational
modificationsThe two subunits are derived from the precursor sequence by an autocatalytic mechanism. -
Cellular localization
Cytoplasm. - Information by UniProt
Associated products
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab276116 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 45 kDa.
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| Notes |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 45 kDa. |
Images
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Western blot - Human CASP1 (Caspase-1) knockout THP-1 cell line (ab276116)All lanes : Anti-Caspase-1 antibody [EPR19672] (ab207802) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate at 40 µg
Lane 2 : CASP1 knockout THP-1 cell lysate at 40 µg
Lane 3 : HeLa cell lysate at 20 µg
Lane 4 : HEK-293 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 35-40, 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Caspase-1 antibody [EPR19672] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab207802 was shown to bind specifically to Caspase-1. A band was observed at 22, 35-40, 48 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP1 knockout cell line ab276116 (knockout cell lysate ab277988). To generate this image, wild-type and CASP1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.False colour image of Western blot: Anti-Caspase-1 antibody [EPR19672] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab207802 was shown to bind specifically to Caspase-1. A band was observed at 22, 35-40, 48 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP1 knockout cell line ab276116 (knockout cell lysate ab277988). To generate this image, wild-type and CASP1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Western blot - Human CASP1 (Caspase-1) knockout THP-1 cell line (ab276116)All lanes : Anti-CASP1 antibody at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate at 40 µg
Lane 2 : THP-1 cell lysate at 40 µg
Lane 3 : HeLa cell lysate at 20 µg
Lane 4 : HEK-293 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 35-40, 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CASP1 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to CASP1. A band was observed at 35-40, 48 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP1 knockout cell line ab276116 (knockout cell lysate ab277988). To generate this image, wild-type and CASP1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab276116 has not yet been referenced specifically in any publications.