Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20885-203] to cleaved C-terminal GSDMD
- Suitable for: WB
- Reacts with: Human
Product nameAnti-cleaved C-terminal GSDMD antibody [EPR20885-203]
See all cleaved C-terminal GSDMD primary antibodies
DescriptionRabbit monoclonal [EPR20885-203] to cleaved C-terminal GSDMD
Tested applicationsSuitable for: WBmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Human
Synthetic peptide within Human cleaved C-terminal GSDMD aa 250-350. The exact sequence is proprietary.
Database link: P57764
- WB: SiHa (treated with 5 µg LPS/10E6 cells by electroporation for 2h.), cell lysate plus concentrated supernatant.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab227821 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 53 kDa).|
FunctionGasdermin-D, N-terminal: Promotes pyroptosis in response to microbial infection and danger signals. Produced by the cleavage of gasdermin-D by inflammatory caspases CASP1 or CASP4 in response to canonical, as well as non-canonical (such as cytosolic LPS) inflammasome activators (PubMed:26375003, PubMed:26375259, PubMed:27418190). After cleavage, moves to the plasma membrane where it strongly binds to inner leaflet lipids, including monophosphorylated phosphatidylinositols, such as phosphatidylinositol 4-phosphate, bisphosphorylated phosphatidylinositols, such as phosphatidylinositol (4,5)-bisphosphate, as well as phosphatidylinositol (3,4,5)-bisphosphate, and more weakly to phosphatidic acid and phosphatidylserine (PubMed:27281216). Homooligomerizes within the membrane and forms pores of 10 - 15 nanometers (nm) of inner diameter, possibly allowing the release of mature IL1B and triggering pyroptosis (PubMed:27418190, PubMed:27281216). Exhibits bactericidal activity. Gasdermin-D, N-terminal released from pyroptotic cells into the extracellular milieu rapidly binds to and kills both Gram-negative and Gram-positive bacteria, without harming neighboring mammalian cells, as it does not disrupt the plasma membrane from the outside due to lipid-binding specificity (PubMed:27281216). Under cell culture conditions, also active against intracellular bacteria, such as Listeria monocytogenes (By similarity). Strongly binds to bacterial and mitochondrial lipids, including cardiolipin. Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine (PubMed:27281216).
Tissue specificityExpressed in the suprabasal cells of esophagus, as well as in the isthmus/neck, pit, and gland of the stomach, suggesting preferential expression in differentiating cells.
Sequence similaritiesBelongs to the gasdermin family.
DomainIntramolecular interactions between N- and C-terminal domains may be important for autoinhibition in the absence of cleavage by inflammatory caspases CASP1 or CASP4. The intrinsic pyroptosis-inducing activity is carried by gasdermin-D, N-terminal, that is released upon cleavage by inflammatory caspases.
modificationsCleavage at Asp-275 by CASP1 (mature and uncleaved precursor forms) or CASP4 relieves autoinhibition and is sufficient to initiate pyroptosis.
Cellular localizationCell membrane. Secreted. Released in the extracellular milieu following pyroptosis and Cytoplasm, cytosol. Inflammasome. In response to a canonical inflammasome stimulus, such as nigericin, recruited to NLRP3 inflammasone with similar kinetics to that of uncleaved CASP1 precursor.
- Information by UniProt
- C-terminal antibody
- Gasdermin domain-containing protein 1 antibody
- Gasdermin-D antibody
All lanes : Anti-cleaved C-terminal GSDMD antibody [EPR20885-203] (ab227821) at 1/1000 dilution
Lane 1 : SiHa (human cervical squamous cell carcinoma), cell lysate plus concentrated cell supernatant
Lane 2 : SiHa (treated with 5 µg LPS/ 10E6 cells by electroporation for 2h.), cell lysate plus concentrated supernatant.
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
Predicted band size: 53 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
The image is kindly provided by Dr Feng Shao.
The molecular weight observed is consistent with literature (PMID: 26375003).
Blocking/Dilution buffer: 5% NFDM/TBST.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab227821 has not yet been referenced specifically in any publications.