Recombinant Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
![Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)](https://www.abcam.com/ps/products/215/ab215203/Images/ab215203-1-anti-cleaved-n-terminal-gsdmd-antibody-epr20829-408-western-blot-a431-human.jpg "Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20829-408] to cleaved N-terminal GSDMD
- Suitable for: WB
- Reacts with: Human
Overview
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Product name
Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] -
Description
Rabbit monoclonal [EPR20829-408] to cleaved N-terminal GSDMD -
Host species
Rabbit -
Specificity
The N-term cleaved Gasdermin-D needs inflammatory caspases in response to canonical and non-canonical inflammasome activators (PubMed: 31548300, PubMed: 27418190, PubMed: 26375259). If need to detect cleavage of GSDMD, please ensure that the samples are treated with inflammation before testing.
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Tested applications
Suitable for: WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human cleaved N-terminal GSDMD aa 250 to the C-terminus. The exact sequence is proprietary. Full length GSDMD contains 484 amino acids, however the immunogen is contained within the N-terminal domain for which the C-terminus is aa 275.
Database link: P57764 -
Positive control
- WB: Salmonella Infected (MOI:100) A431 whole cell lysate; THP-1 treated with 500 ng/ml EprI for 2h, whole cell lysate.
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General notes
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20829-408 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
Our Abpromise guarantee covers the use of ab215203 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/1000. Detects a band of approximately 31 kDa (predicted molecular weight: 53 kDa). |
Target
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Function
Gasdermin-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 specificity
Expressed 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 similarities
Belongs to the gasdermin family. -
Domain
Intramolecular 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. -
Post-translational
modificationsCleavage at Asp-275 by CASP1 (mature and uncleaved precursor forms) or CASP4 relieves autoinhibition and is sufficient to initiate pyroptosis. -
Cellular localization
Cell 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
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Database links
- Entrez Gene: 79792 Human
- SwissProt: P57764 Human
- Unigene: 118983 Human
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Alternative names
- C-terminal antibody
- Gasdermin domain-containing protein 1 antibody
- Gasdermin-D antibody
see all
Images
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Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203) at 1/1000 dilution + Salmonella Infected (MOI:100) A431 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesLysate was kindly provided by Dr Feng Shao.
The molecular weight observed is consistent with literature (PMID: 26375003).
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)](https://www.abcam.com/ps/products/215/ab215203/Images/ab215203-2-anti-cleaved-n-terminal-gsdmd-antibody-epr20829-408-western-blot-thp1-human.jpg)
Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)All lanes : Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203) at 1/1000 dilution
Lane 1 : Untreated THP-1 (human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : THP-1 treated with 500 ng/ml EprI for 2h, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesLysate was kindly provided by Dr Feng Shao.
The molecular weight observed is consistent with literature (PMID: 26375003).
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)](https://www.abcam.com/ps/products/215/ab215203/Images/ab215203-3-benefits-of-recombinant-antibodies.png)
Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
Protocols
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.
References (3)
ab215203 has been referenced in 3 publications.
- Wang K et al. Structural Mechanism for GSDMD Targeting by Autoprocessed Caspases in Pyroptosis. Cell N/A:N/A (2020). PubMed: 32109412
- Tan YF et al. Inhibition of BRD4 prevents proliferation and epithelial-mesenchymal transition in renal cell carcinoma via NLRP3 inflammasome-induced pyroptosis. Cell Death Dis 11:239 (2020). PubMed: 32303673
- Zhao Q et al. Novel HSP90-PI3K Dual Inhibitor Suppresses Melanoma Cell Proliferation by Interfering with HSP90-EGFR Interaction and Downstream Signaling Pathways. Int J Mol Sci 21:N/A (2020). PubMed: 32156008
