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AB275493

Apoptosis pathway knockout cell lines panel

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FAS KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

View Alternative Names

ALPS 1A, ALPS2B, APO 1, APO 1 cell surface antigen, APT-1, Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein, Apo-1 antigen, Apoptosis antigen 1, Apoptosis regulator BAX, Apoptosis related cysteine peptidase, Apoptosis-mediating surface antigen FAS, Apoptotic cysteine protease, Apoptotic protease Mch-5, BAXA, BAX_HUMAN, BCL2 associated X protein, BCL2 associated X protein omega, BCL2 associated X protein transcript variant delta2, BING 2, Bax-protein, Baxdelta2G9, Baxdelta2G9omega, Baxdelta2omega, Bcl-2-like protein 4, Bcl2-L-4, CAP 4, CARD 3, CARD carrying kinase, CARD containing ICE associated kinase, CARD containing interleukin 1 beta converting enzyme (ICE) associated kinase, CARD-containing IL-1 beta ICE-kinase, CARD-containing interleukin-1 beta-converting enzyme-associated kinase, CARDIAK, CASP8_HUMAN, CD 95, CD 95 antigen, CED 3, CENP-C binding protein, CLARP kinase, Caspase 8, Caspase 8 apoptosis related cysteine peptidase, Caspase IIX, Caspase-8 subunit p10, DAP 6, DAXX_HUMAN, Death associated protein 6, Death domain associated protein, Death domain-associated protein 6, Delta Fas, Delta Fas/APO 1/CD95, EAP 1, ETS1-associated protein 1, FADD protein, FADD-homologous ICE/CED-3-like protease, FADD-like ICE, FADD_HUMAN, FAS 1, FAS 827dupA, FAS Antigen, FAS-associated death domain protein, FAS-associating death domain-containing protein, FASLG receptor, FASTM, FLICE, FLJ17672, Fas (TNF receptor superfamily, member 6), Fas (TNFRSF6) associated via death domain, Fas AMA, Fas TNFRSF6 associated via death domain, Fas associated via death domain, Fas associating protein, Fas associating protein with death domain, Fas binding protein, Fas cell surface death receptor, Fas death domain-associated protein, GIG 3, GIG 30, Growth inhibiting gene 30, Growth-inhibiting gene 3 protein, H sapiens mRNA for mediator of receptor induced toxicity, ICE-like apoptotic protease 5, MACH, MACH alpha 1/2/3 protein, MACH beta 1/2/3/4 protein, MACH5, MCH 5, MGC126245, MGC126246, MGC78473, MGC8528, MORT 1, MORT1-associated CED-3 homolog, Mediator of receptor induced toxicity, OTTHUMP00000163717, OTTHUMP00000163720, OTTHUMP00000163724, OTTHUMP00000163725, OTTHUMP00000165062, OTTHUMP00000165063, OTTHUMP00000165064, OTTHUMP00000206552, OTTHUMP00000206582, Protein FADD, RICK, RIP-like-interacting CLARP kinase, RIPK2_HUMAN, Receptor interacting protein (RIP) like interacting caspase like apoptosis regulatory protein (CLARP) kinase, Receptor interacting serine threonine kinase 2, Receptor-interacting protein 2, Receptor-interacting serine/threonine-protein kinase 2, Surface antigen APO1, TNF receptor superfamily, member 6, TNFRSF, TNFRSF 6, TNR6_HUMAN, Tumor necrosis factor receptor superfamily member 6, Tyrosine-protein kinase RIPK2, UNQ277/PRO314/PRO34092, hDaxx, membrane isoform alpha, sFAS

9 Images
Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : CASP8 knockout HeLa cell lysate (20μg)
Lane 3 : Jurkat cell lysate (20μg)
Lane 4 : SH-SY5Y cell lysate (20μg)
Lanes 1- 4 : Merged signal (red and green). Green - ab32125 observed at 55 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab32125 Anti-Caspase-8 antibody [E6] was shown to specifically react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264958 (knockout cell lysate ab256857) was used. Wild-type and Caspase-8 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32125 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 3000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : BAX knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab182734 observed at 21 kDa. Red - loading control ab8245 observed at 37 kDa.
ab182734 Recombinant Anti-Bax antibody [EPR18284 was shown to specifically react with BAX in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type and BAX knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : FAS knockout HeLa cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab133619 observed at 37 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133619 Anti-Fas antibody [EPR5700] was shown to specifically react with Fas in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265260 (knockout cell lysate ab256911) was used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : CASP8 knockout HeLa cell lysate (20μg)
Lane 3 : Jurkat cell lysate (20μg)
Lane 4 : SH-SY5Y cell lysate (20μg)
Lanes 1- 4 : Merged signal (red and green). Green - ab32397 observed at 55 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab32397 Rabbit monoclonal [EPR2418Y] to IRF3 was shown to specifically react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264958 (knockout cell lysate ab256857) was used. Wild-type and Caspase-8 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : RIPK2 knockout HeLa cell lysate (20 μg)

Lane 3 : Ramos cell lysate (20 μg)

Lane 4 : Jurkat cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab75257 observed at 65 kDa. Red - loading control ab181602 observed at 37 kDa.

ab75257 Anti-RIP2 antibody [AF28D3] was shown to specifically react with RIP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264688 (knockout cell lysate ab258636) was used. Wild-type and RIP2 knockout samples were subjected to SDS-PAGE. ab75257 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : FADD knockout HeLa cell lysate (20 μg)

Lane 3 : A431 cell lysate (20 μg)

Lane 4 : Jurkat cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab119059 observed at 25 kDa. Red - loading control ab181602 observed at 37 kDa.

ab119059 Anti-FADD antibody [OTI1C11] was shown to specifically react with FADD in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261817 (knockout cell lysate ab257261) was used. Wild-type and FADD knockout samples were subjected to SDS-PAGE. ab119059 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : FADD knockout HeLa cell lysate (20 μg)

Lane 3 : A431 cell lysate (20 μg)

Lane 4 : Jurkat cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab108601 observed at 25 kDa. Red - loading control ab8245 observed at 37 kDa.

ab108601 Anti-FADD antibody [EPR4415] was shown to specifically react with FADD in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261817 (knockout cell lysate ab257261) was used. Wild-type and FADD knockout samples were subjected to SDS-PAGE. ab108601 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : BAX knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab32503 observed at 21 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32503 Recombinant Anti-Bax antibody [E63] was shown to specifically react with BAX in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type and BAX knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32503 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

false

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)
  • WB

Lab

Western blot - Apoptosis pathway knockout cell lines panel (AB275493)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : DAXX knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab32140 observed at 100 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32140 Recombinant Anti-Daxx antibody [E94] was shown to specifically react with Daxx in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265233 (knockout cell lysate ab257408) was used. Wild-type and Daxx knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32140 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

Disease

Adenocarcinoma

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Reactivity data

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Product details

A collection of 6 knockout cell lines involved in the apoptotic cell death pathways, in one kit for your convenience, with matching parental cell lines.

Find out more about our apoptosis assays.

Included in this panel:

FAS knockout HeLa cell Line (ab265260) + recommended control (ab255928)

FADD knockout HeLa cell Line (ab261817) + recommended control (ab255928)

DAXX knockout HeLa cell Line (ab265233) + recommended control (ab255928)

CASP8 knockout HeLa cell Line (ab264958) + recommended control (ab255448)

BAX knockout HeLa cell Line (ab255363) + recommended control (ab255448)

RIPK2 (RIP2) knockout HeLa cell Line (ab264688) + recommended control (ab255448)

Parts of the panels can be purchased separately. For further details please contact technical@abcam.com.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
FAS
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

FADD Bax Daxx Fas RIP2 and Caspase-8 are vital components in cell death processes. Mechanically these proteins interact to regulate apoptosis necroptosis and cellular stress responses. FADD (Fas-Associated protein with Death Domain) is a 23-kDa cytoplasmic adaptor protein actively engaged in transducing apoptotic signals. Caspase-8 often associated with initiator caspases exists primarily in the cytoplasm with an approximate mass of 55 kDa and initiates apoptosis by cleaving downstream effector caspases. Fas a cell surface receptor sometimes called CD95 triggers apoptosis upon ligand binding. Bax a pro-apoptotic member of Bcl-2 family facilitates mitochondrial membrane permeabilization. Daxx or Death-domain associated protein can shuttle between the nucleus and cytoplasm influencing apoptosis and transcription regulation. RIP2 (also known as RIPK2) is a serine/threonine-protein kinase that contributes to innate immune responses.
Biological function summary

FADD Bax Daxx Fas RIP2 and Caspase-8 play key roles in mediating apoptotic pathways. FADD forms complexes with death receptors like Fas to initiate apoptotic signaling cascades. Daxx reportedly binds death receptors modulating apoptotic activities. Caspase-8 freely catalyzes the activation of downstream caspase enzymes effectively driving the cell toward apoptotic execution. Bax integration into mitochondrial pathways disrupts membrane potential releasing cytochrome c that further supports caspase-9 activation. RIP2 while known for immune responses also synergizes with these apoptotic players in certain contexts. These proteins collectively orchestrate a balance of cell survival and death essential for normal cellular homeostasis and tissue development.

Pathways

FADD Bax Daxx Fas RIP2 and Caspase-8 are fundamental to the extrinsic apoptosis and necroptosis pathways. In the extrinsic pathway Fas ligation triggers FADD recruitment which then binds with death effector domain to activate Caspase-8 facilitating the apoptotic cascade. The involvement of Bax influences the mitochondrial apoptosis pathway by promoting cytochrome c release following intrinsic apoptotic signals. RIP2 though more common in inflammatory pathways may cross-interact with these proteins in the context of cellular stress responses. These interactions highlight a tight regulatory network with precise cross-communication between apoptotic and survival signals.

Disrupted regulation of FADD Bax Daxx Fas RIP2 and Caspase-8 connects significantly with cancer and autoimmune diseases. Deregulated Fas-mediated apoptosis often appears in various cancer forms where deficient apoptosis contributes to uncontrolled cell proliferation. Caspase-8 mutations or impaired expression link to neurodegenerative diseases and some cancers due to unregulated cell death. In autoimmune diseases like systemic lupus erythematosus abnormal expression of Fas and related apoptotic proteins like Daxx may participate in perpetuating an immune attack on self-cells. The core connection of these proteins to apoptosis and immune regulation underlines their potential as therapeutic targets in these pathological states.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

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