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AB276122

Human CASP9 knockout THP-1 cell line

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CASP9 KO cell line available to order. KO validated by. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

APAF-3, Apoptosis related cysteine peptidase, Apoptotic protease Mch-6, Apoptotic protease-activating factor 3, CASP9_HUMAN, Caspase 9 Dominant Negative, Caspase 9 apoptosis related cysteine peptidase, Caspase 9c, Caspase-9, Caspase-9 subunit p10, ICE-LAP6, ICE-like apoptotic protease 6, MCH6, PPP1R56, RNCASP9, protein phosphatase 1, regulatory subunit 56

3 Images
Western blot - Human CASP9 knockout THP-1 cell line (AB276122)
  • WB

Lab

Western blot - Human CASP9 knockout THP-1 cell line (AB276122)

False colour image of Western blot : Anti-Caspase-9 antibody [EPR18108] staining at 1/1000 dilution shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab185719 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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.

All lanes:

Western blot - Anti-Caspase-9 antibody [EPR18108] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-epr18108-ab185719'>ab185719</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

CASP9 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human CASP9 knockout THP-1 cell line (ab276122)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 45 kDa

false

Western blot - Human CASP9 knockout THP-1 cell line (AB276122)
  • WB

Lab

Western blot - Human CASP9 knockout THP-1 cell line (AB276122)

False colour image of Western blot : Anti-Caspase-9 antibody [EPR18107] staining at 1/2000 dilution shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab202068 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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.

All lanes:

Western blot - Anti-Caspase-9 antibody [EPR18107] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-epr18107-ab202068'>ab202068</a>) at 1/2000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

CASP9 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human CASP9 knockout THP-1 cell line (ab276122)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 45 kDa

false

Western blot - Human CASP9 knockout THP-1 cell line (AB276122)
  • WB

Lab

Western blot - Human CASP9 knockout THP-1 cell line (AB276122)

False colour image of Western blot : Anti-Caspase-9 antibody [E23] staining at 1/1000 dilution shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab32539 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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.

All lanes:

Western blot - Anti-Caspase-9 antibody [E23] (<a href='/en-us/products/primary-antibodies/caspase-9-antibody-e23-ab32539'>ab32539</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

CASP9 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human CASP9 knockout THP-1 cell line (ab276122)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 45 kDa

false

Key facts

Cell type

THP-1

Species or organism

Human

Tissue

Blood

Form

Liquid

form

Disease

Acute Monocytic Leukemia

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?

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Properties and storage information

Gene name
CASP9
Gene editing type
Knockout
Gene editing method
CRISPR technology
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

Product protocols

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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