- PA5-29342 - Invitrogen Antibodies CASP1 antibody

Submitted references Diacerein counteracts acetaminophen-induced hepatotoxicity in mice via targeting NLRP3/caspase-1/IL-1β and IL-4/MCP-1 signaling pathways.

Elshal M, Abdelmageed ME
Archives of pharmacal research 2022 Mar;45(3):142-158

A promiscuous inflammasome sparks replication of a common tumor virus.

Burton EM, Goldbach-Mansky R, Bhaduri-McIntosh S
Proceedings of the National Academy of Sciences of the United States of America 2020 Jan 21;117(3):1722-1730

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using Caspase 1 Polyclonal Antibody (Product # PA5-29342). Sample (30 µg of whole cell lysate). Lane A: A549. 10% SDS PAGE. Caspase 1 Polyclonal Antibody (Product # PA5-29342) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockout of CASP1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR917229_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CASP1 was performed by loading 30 µg of HT-29 wild type (Lane 1), HT-29 wild type treated with 10 ng/mL IFN gamma for 24hrs (Lane 2),HT-29 Cas9 (Lane 3), HT-29 Cas9 treated with 10 ng/mL IFN gamma for 24hrs (Lane 4), HT-29 CASP1 KO (Lane 5) and HT-29 CASP1 KO treated with 10 ng/mL IFN gamma for 24hrs (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein 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 Anti-Caspase 1 Polyclonal Antibody (Product # PA5-29342, 1:3000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CASP1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Caspase 1 Polyclonal Antibody(Product # PA5-29342) and a 48kDa band corresponding to Caspase 1 was observed across cell lines tested except MCF-7, SH-SY5Y and HEK-293 which is reported to be low. Whole Cell Extract-WCL (30 µg lysate) of THP-1 (Lane 1), NK-92 (Lane 2), MCF7 (Lane 3), SH-SY5Y (Lane 4) and HEK-293 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Caspase 1 in methanol-fixed HeLa cells using a Caspase 1 polyclonal antibody (Product # PA5-29342) at a 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Caspase 1 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 45 minutes at room temperature. The cells were labeled with Caspase 1 Polyclonal Antibody (Product # PA5-29342) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Effect of diacerein (DCN) on A NLR family pyrin domain containing 3 (NLRP3) and B Interlukin-1 beta (IL-1beta) levels in hepatic tissue of acetaminophen-injected mice. Data are expressed as mean +- SEM (n = 8). #p < 0.05, ##p < 0.01 compared to control group, *p < 0.05, **p < 0.01 compared to acetaminophen-injected group, Fp < 0.05, FFp < 0.01 compared to DCN 4 h prior to acetaminophen group, *p < 0.05, **p < 0.01 compared to DCN 2 h post-acetaminophen group using one-way ANOVA followed by Tukey-Kramer multiple comparisons post hoc test. C Effect of diacerein (DCN) on hepatic caspase-1 assessed by immunohistochemistry (X 400, bar = 50 um); (a) hepatic sections of control group showing negative staining; (b) hepatic sections of DCN group showing negative staining; (c) hepatic sections of acetaminophen group showing marked positive brown cytoplasmic expression in hepatocytes (black arrows) associated with area of lesions around central veins; (d) hepatic sections of DCN 4 h prior to acetaminophen group showing a significant reduction of caspase-1 cytoplasmic expression (black arrows); (e) hepatic sections of DCN 2 h post-acetaminophen group showing a significant reduction of caspase-1 cytoplasmic expression (black arrows), and (f) scatter dot plots of the immunohistopathological assessment of hepatic caspase-1 scores. $p < 0.05, $$p < 0.01 compared to control group, +p < 0.05, ++p < 0.01 compared to compared to acetaminophen group using Kruskal-Wallis followed by Dunn's Multipl

PA5-29342

Antibody data