MA5-56476

antibody from Invitrogen Antibodies

Targeting: CASP8 Casp-8, FLICE, MACH, MCH5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-56476

Provider

Invitrogen Antibodies

Product name

Caspase 8 Recombinant Rabbit Monoclonal Antibody (PSH04-77)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

PSH04-77

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Caspase 8 in Lane 1: HAP1-parental cell lysate, Lane 2: HAP1-Caspase-8 KD cell lysate. (Lysates/Proteins at 10 µg/Lane). Samples were incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) using a dilution of 1:1,000 Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST (1 hr, room temperature). Followed by using Goat Anti-Rabbit IgG-HRP Secondary Antibody at dilution 1:50,000 (1hr, room temp). Predicted band size: 55 kDa; Observed band size: 54,55 kDa; Exposure time: 180 seconds.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Caspase 8 in Lane 1: Jurkat cell lysate, Lane 2: Jurkat treated with 25μM Etoposide for 5 hours cell lysate, Lane 3: Jurkat treated with 25μM Etoposide for 16 hours cell lysate, Lane 4: Jurkat treated with 1μM Staurosporine for 3 hours cell lysate, Lane 5: HeLa cell lysate, Lane 6: HeLa treated with 1μM Staurosporine for 3 hours cell lysate, Lane 7: HeLa treated with 100μM Etoposide for 4 hours cell lysate. (Lysates/Proteins at 30 µg/Lane). Samples were incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) using a dilution of 1:2,000 Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST (1 hr, room temperature). Followed by using Goat Anti-Rabbit IgG-HRP Secondary Antibody at dilution 1:50,000 (1hr, room temp). Predicted band size: 55 kDa; Observed band size: 18-55 kDa; Exposure time: 3 minutes.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of Caspase 8 with HeLa cells . Cells were fixed in 4% paraformaldehyde (20 min, room temp), permeabilized with 0.1% Triton X-100 in PBS (5 min, room temp), then blocked with 1% BSA in 10% negative goat serum (1 hr, room temp). Next they were incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476)at a dilution of 1:100 in 1% BSA in PBST overnight at 4 ℃. Followed by Goat Anti-Rabbit IgG H&L at a dilution of 1:1,000. DAPI was used to stain the cell nuclei (blue).

Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Caspase 8 was immunoprecipitated from 0.2 mg HeLa treated with 1μM Staurosporine for 3 hours cell lysate with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) at 1:1,000 dilution. Followed by Anti-Rabbit IgG for IP Nano-secondary antibody at 1:5,000 dilution (1 hr, room temperature). Lane 1: HeLa treated with 1μM Staurosporine for 3 hours cell lysate; Lane 2: (MA5-56476) IP in HeLa treated with 1μM Staurosporine for 3 hours cell lysate; Lane 3: Rabbit IgG in HeLa treated with 1μM Staurosporine for 3 hours cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 1 minute 28 seconds.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Caspase 8 with paraffin-embedded human tonsil tissue. The tissues were blocked in 1% BSA (20 min, room temp), washed with ddH2O and PBS, incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) at a dilution of 1:1,000 (1 hr, room temp).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Caspase 8 with paraffin-embedded human liver tissue. The tissues were blocked in 1% BSA (20 min, room temp), washed with ddH2O and PBS, incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) at a dilution of 1:200 (1 hr, room temp).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Caspase 8 with HeLa cells. The cells were fixed, permeabilized and incubated with Caspase 8 Recombinant Monoclonal Antibody (Product # MA5-56476) at a dilution of 1μg/mL (red) and compared with Rabbit IgG Isotype Control (green). Followed by iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1:1,000 dilution (30 min, 4℃).