Anti-Caspase-7 antibody [EPR17029]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [EPR17029] (AB181579)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling Caspase-7 with ab181579 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on NIH/3T3 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows :
1. ab181579 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

Please see Pubmed IDs : 22464733 and 11829465 for further information on Caspase-7 localization.

• IP

Supplier Data

Immunoprecipitation - Anti-Caspase-7 antibody [EPR17029] (AB181579)

Caspase-7 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) with ab181579 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab181579 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : NIH/3T3 whole cell extract. Lane 2 : PBS instead of extract.

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed band at 35kDa band is the pro-caspase-7.

All lanes:

Immunoprecipitation - Anti-Caspase-7 antibody [EPR17029] (ab181579)

Predicted band size: 34 kDa

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• WB

Supplier Data

Western blot - Anti-Caspase-7 antibody [EPR17029] (AB181579)

Observed band size : The 35kDa band is the pro-caspase 7; 32kDa band is the N-terminal propeptide cleaved caspase 7; the 27kDa band is the truncated form and the 11kDa band is Caspase 7 subunit p11.
Lanes 1-4 were from the same blot; the exposure time for lanes 1-3 is 1 min and for lane 4 is 2 min.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Caspase-7 antibody [EPR17029] (ab181579) at 1/5000 dilution

Lane 1:

Rat brain tissue lysate at 10 µg

Lane 2:

Rat heart tissue lysate at 10 µg

Lane 3:

Rat kidney tissue lysate at 10 µg

Lane 4:

Rat spleen tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 34 kDa

false

• WB

Supplier Data

Western blot - Anti-Caspase-7 antibody [EPR17029] (AB181579)

Observed band size : 35kDa band is the pro-Caspase 7, 32kDa band is the N-terminal propeptide cleaved Caspase 7, and the 27kDa band is an N-terminal truncated caspase 7.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Caspase-7 antibody [EPR17029] (ab181579) at 1/5000 dilution

Lane 1:

Mouse heart tissue lysate at 10 µg

Lane 2:

Mouse kidney tissue lysate at 10 µg

Lane 3:

Mouse spleen tissue lysate at 10 µg

Lane 4:

C6 (Rat glial tumor cells) lysate at 10 µg

Lane 5:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate at 10 µg

Lane 6:

PC-12 (Rat adrenal gland pheochromocytoma) lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embyro fibroblast cells) lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 34 kDa

Observed band size: 27 kDa,32 kDa,35 kDa

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