710431

antibody from Invitrogen Antibodies

Targeting: CASP3 apopain, CPP32, CPP32B, Yama

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

710431

Provider

Invitrogen Antibodies

Product name

Caspase 3 Recombinant Polyclonal Antibody (22HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

22HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

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

References

Submitted references

The inhibition of histone deacetylase 8 suppresses proliferation and inhibits apoptosis in gastric adenocarcinoma.

Song S, Wang Y, Xu P, Yang R, Ma Z, Liang S, Zhang G
International journal of oncology 2015 Nov;47(5):1819-28

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Caspase-3 in whole cell extracts from HeLa, HeLa treated with Staurosporine (1uM, 3 hrs), Jurkat, and Jurkat treated with Staurosporine (1uM, 3 hrs) (lanes 1- 4 respectively) using a Caspase-3 Recombinant Rabbit Polyclonal Antibody (Product # 710431) at a dilution of 2 µg/mL. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~35kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Caspase-3 was performed by loading 30 µg of HeLa, HeLa treated with Staurosporine (3 uM for 1 hour), Jurkat and Jurkat treated with Staurosporine (3 uM for 1 hour) using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. Caspase-3 was detected at ~35 kDa using Caspase-3 Recombinant Rabbit Polyclonal Antibody (Product # 710431) at a 1:500 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Caspase-3 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Caspase-3 Recombinant Rabbit Polyclonal Antibody (Product # 710431) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381) Panels d is a merged image showing cytoplasmic localization and panel e a is no primary antibody control. The images were captured using a Nikon microscope at 20X magnification.