MA5-26748

antibody from Invitrogen Antibodies

Targeting: CASP4 ICE(rel)II, ICH-2, TX

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-26748

Provider

Invitrogen Antibodies

Product name

Caspase 4 Monoclonal Antibody (OTI4A2)

Antibody type

Monoclonal

Antigen

Recombinant protein fragment

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

OTI4A2

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Caspase 4 Monoclonal Antibody (Product # MA5-26748) and a 45 kDa, 36 kDa bands corresponding to Caspase 4 were observed across cell lines tested and increased upon PMA and LPS treatment. Whole cell lysate extracts (30 µg lysate) of THP-1 (Lane 1) and THP-1 treated with PMA (80nM) and LPS (10ng/mL) (Lane 2) simultaneously for 16 hrs and 5 hrs were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis Caspase 4 Monoclonal Antibody was performed using 70% confluent log phase THP-1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled Caspase 4 Monoclonal antibody (Product MA5-26748) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175) 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 nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.