JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB256303

Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free

Be the first to review this product! Submit a review

|

(1 Publication)

Knockout Tested Rabbit Recombinant Monoclonal Caspase-4 antibody. Carrier free. Suitable for WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 1 publication.

View Alternative Names

ICH2, CASP4, Caspase-4, CASP-4, ICE and Ced-3 homolog 2, ICE(rel)-II, Mih1, Protease TX, ICH-2

3 Images
Flow Cytometry (Intracellular) - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)
  • Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized IM-9 (human multiple myeloma B Lymphoblast) cell line labeling Caspase 4 with ab238124 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150097), at 1/5000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238124).

Flow Cytometry (Intracellular) - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)
  • Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)

Intracellular flow cytometric analysis of THP-1 (Human monocytic leukemia monocyte) cells labeling Caspase-4 with ab238124 at 1/60 (right panel, red) compared with HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells (left panel, red). Goat anti rabbit IgG (Alexa Fluor® 488, ab150097), at 1/5000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Negative control : HEK-293T (PMID : 25119034).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238124).

Western blot - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)
  • WB

Lab

Western blot - Anti-Caspase-4 antibody [EPR20921-83] - BSA and Azide free (AB256303)

This data was developed using the same antibody clone in a different buffer formulation (ab238124).

Western blot : Anti-CASP4 antibody [EPR20921-83] (ab238124) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab238124 was shown to bind specifically to CASP4. A band was observed at 43 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CASP4 knockout cell line. To generate this image, wild-type and CASP4 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Caspase-4 antibody [EPR20921-83] (<a href='/en-us/products/primary-antibodies/caspase-4-antibody-epr20921-83-ab238124'>ab238124</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

CASP4 knockout HCT 116 cell lysate at 20 µg

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 43 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR20921-83

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>" } } }

Product details

ab256303 is the carrier-free version of ab238124.

This antibody was developed in collaboration with Dr Feng Shao's lab, NIBS.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

Inflammatory caspase that acts as the effector of the non-canonical inflammasome by mediating lipopolysaccharide (LPS)-induced pyroptosis (PubMed : 25119034, PubMed : 26375003, PubMed : 32109412, PubMed : 34671164, PubMed : 37001519, PubMed : 37993712, PubMed : 37993714). Also indirectly activates the NLRP3 and NLRP6 inflammasomes (PubMed : 23516580, PubMed : 26375003, PubMed : 32109412, PubMed : 7797510). Acts as a thiol protease that cleaves a tetrapeptide after an Asp residue at position P1 : catalyzes cleavage of CGAS, GSDMD and IL18 (PubMed : 15326478, PubMed : 23516580, PubMed : 26375003, PubMed : 28314590, PubMed : 32109412, PubMed : 37993712, PubMed : 37993714, PubMed : 7797510). Effector of the non-canonical inflammasome independently of NLRP3 inflammasome and CASP1 : the non-canonical inflammasome promotes pyroptosis through GSDMD cleavage without involving secretion of cytokine IL1B (PubMed : 25119034, PubMed : 25121752, PubMed : 26375003, PubMed : 31268602, PubMed : 32109412, PubMed : 37993712, PubMed : 37993714). In the non-canonical inflammasome, CASP4 is activated by direct binding to the lipid A moiety of LPS without the need of an upstream sensor (PubMed : 25119034, PubMed : 25121752, PubMed : 29520027, PubMed : 32510692, PubMed : 32581219, PubMed : 37993712). LPS-binding promotes CASP4 activation and CASP4-mediated cleavage of GSDMD and IL18, followed by IL18 secretion through the GSDMD pore, pyroptosis of infected cells and their extrusion into the gut lumen (PubMed : 25119034, PubMed : 25121752, PubMed : 37993712, PubMed : 37993714). Also indirectly promotes secretion of mature cytokines (IL1A and HMGB1) downstream of GSDMD-mediated pyroptosis via activation of the NLRP3 and NLRP6 inflammasomes (PubMed : 26375003, PubMed : 32109412). Involved in NLRP3-dependent CASP1 activation and IL1B secretion in response to non-canonical activators, such as UVB radiation or cholera enterotoxin (PubMed : 22246630, PubMed : 23516580, PubMed : 24879791, PubMed : 25964352, PubMed : 26173988, PubMed : 26174085, PubMed : 26508369). Involved in NLRP6 inflammasome-dependent activation in response to lipoteichoic acid (LTA), a cell-wall component of Gram-positive bacteria, which leads to CASP1 activation and IL1B secretion (PubMed : 33377178). Involved in LPS-induced IL6 secretion; this activity may not require caspase enzymatic activity (PubMed : 26508369). The non-canonical inflammasome is required for innate immunity to cytosolic, but not vacuolar, bacteria (By similarity). Plays a crucial role in the restriction of S.typhimurium replication in colonic epithelial cells during infection (PubMed : 25121752, PubMed : 25964352). Activation of the non-canonical inflammasome in brain endothelial cells can lead to excessive pyroptosis, leading to blood-brain barrier breakdown (By similarity). Pyroptosis limits bacterial replication, while cytokine secretion promotes the recruitment and activation of immune cells and triggers mucosal inflammation (PubMed : 25121752, PubMed : 25964352, PubMed : 26375003). May also act as an activator of adaptive immunity in dendritic cells, following activation by oxidized phospholipid 1-palmitoyl-2-arachidonoyl- sn-glycero-3-phosphorylcholine, an oxidized phospholipid (oxPAPC) (By similarity). Involved in cell death induced by endoplasmic reticulum stress and by treatment with cytotoxic APP peptides found in Alzheimer's patient brains (PubMed : 15123740, PubMed : 22246630, PubMed : 23661706). Cleavage of GSDMD is not strictly dependent on the consensus cleavage site but depends on an exosite interface on CASP4 that recognizes and binds the Gasdermin-D, C-terminal (GSDMD-CT) part (PubMed : 32109412). Catalyzes cleavage and maturation of IL18; IL18 processing also depends of the exosite interface on CASP4 (PubMed : 15326478, PubMed : 37993712, PubMed : 37993714). In contrast, it does not directly process IL1B (PubMed : 7743998, PubMed : 7797510, PubMed : 7797592). During non-canonical inflammasome activation, cuts CGAS and may play a role in the regulation of antiviral innate immune activation (PubMed : 28314590).. (Microbial infection) In response to the Td92 surface protein of the periodontal pathogen T.denticola, activated by cathepsin CTSG which leads to production and secretion of IL1A and pyroptosis of gingival fibroblasts.
See full target information CASP4

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Frontiers in cardiovascular medicine 9:1013262 PubMed36684586

2023

SARS-CoV-2 Spike protein activates TMEM16F-mediated platelet procoagulant activity.

Applications

Unspecified application

Species

Unspecified reactive species

Ambra Cappelletto,Harriet E Allan,Marilena Crescente,Edoardo Schneider,Rossana Bussani,Hashim Ali,Ilaria Secco,Simone Vodret,Roberto Simeone,Luca Mascaretti,Serena Zacchigna,Timothy D Warner,Mauro Giacca
View all publications

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com