PA1-26434

antibody from Invitrogen Antibodies

Targeting: CASP9 APAF-3, ICE-LAP6, MCH6, PPP1R56

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

PA1-26434

Provider

Invitrogen Antibodies

Product name

Caspase 9 (active) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

50 µg

Concentration

0.2 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 9 (active) was performed by loading (1) untreated and (2) Etoposide treated Jurkat cell lysates. Proteins were transferred to a membrane and probed with a Caspase 9 (active) Polyclonal Antibody (Product # PA1-26434).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with Staurosporine (1uM for 3hrs) (Lane 2), Jurkat (Lane 3) and Jurkat treated with Staurosporine (1uM for 3hrs) (Lane 4). The blot was probed with Anti-Caspase 9 (active) Polyclonal Antibody (Product # PA1-26434, 1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 37 kDa band corresponding to cleaved Caspase 9 was observed only in the Staurosporine treated cell lines tested. A non-specific band was also observed at ~58 kDa in all the lanes.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Caspase 9 was performed using 70% confluent log phase HeLa cells treated with 1µM of Staurosporine for 3 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Caspase 9 Rabbit Polyclonal Antibody (Product # PA1-26434) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows untreated cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.