Anti-GSDMD antibody

Cat# NB-22-46805-100

Size : 100μL

Brand : Neo Biotech

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Phone : +1 850 650 7790

  • Western blot analysis of extracts of various cell lines, using GSDMD antibody at 1:1000 dilution. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 30s.
  • Immunohistochemistry of paraffin-embedded rat spleen using GSDMD rabbit polyclonal antibody at dilution of 1:100 (40x lens). Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry of paraffin-embedded human colon carcinoma using GSDMD (Full Length+N terminal) rabbit polyclonal antibody at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry of paraffin-embedded human tonsil using GSDMD (Full Length+N terminal) rabbit polyclonal antibody at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of BALB-3T3 cells using GSDMD rabbit polyclonal antibody at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
  • Immunofluorescence analysis of C6 cells using GSDMD rabbit polyclonal antibody at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
  • Immunofluorescence analysis of U-2 OS cells using GSDMD rabbit polyclonal antibody at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
  • Immunoprecipitation analysis of 300ug extracts of Jurkat cells using 3ug GSDMD antibody . Western blot was performed from the immunoprecipitate using GSDMD antibody at a dilition of 1:500.

General Info

Host: Rabbit
Applications: WB/IHC/IF/IP
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-GSDMD (346-484) is suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence and Immunoprecipitation research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.05% Proclin300, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:100-1:500
IHC-P 1:50-1:200
IF/ICC 1:50-1:200
IP 1:100-1:500
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.

Information

Gene Symbol: GSDMD
Gene ID: 79792
Uniprot ID: GSDMD_HUMAN
Immunogen Region: 346-484
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 346-484 of human GSDMD (NP_079012.3).
Immunogen Sequence: DGPAGAVLECLVLSSGMLVP ELAIPVVYLLGALTMLSETQ HKLLAEALESQTLLGPLELV GSLLEQSAPWQERSTMSLPP GLLGNSWGEGAPAWVLLDEC GLELGEDTPHVCWEPQAQGR MCALYASLALLSGLSQEPH

Description

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.
Post Translational Modifications Cleavage at Asp-275 by CASP1 (mature and uncleaved precursor forms), CASP4, CASP5 or CASP8 relieves autoinhibition and is sufficient to initiate pyroptosis. Cleavage by CASP1 and CASP4 is not strictly dependent on the consensus cleavage site on GSDMD but depends on an exosite interface on CASP1 that recognizes and binds the Gasdermin-D, C-terminal (GSDMD-CT) part. Cleavage by CASP8 takes place following inactivation of MAP3K7/TAK1 by Yersinia toxin YopJ. Cleavage at Asp-87 by CASP3 or CAPS7 inactivates the ability to mediate pyroptosis, but generates the Gasdermin-D, p13 chain, which translocates to the nucleus and acts as a transcription regulator. Cleavage by papain allergen generates the Gasdermin-D, p40 chain. Gasdermin-D: Succination of Cys-191 by the Krebs cycle intermediate fumarate, which leads to S-(2-succinyl)cysteine residues, inhibits processing by caspases, and ability to initiate pyroptosis. Succination modification is catalyzed by a non-enzymatic reaction caused by an accumulation of fumarate. (Microbial infection) Cleaved and inactivated by Protease 3C from Human enterovirus 71 (EV71), preventing GSDMD-mediated pyroptosis. (Microbial infection) Cleaved and inactivated by the 3C-like proteinase nsp5 from human coronavirus SARS-CoV-2, preventing GSDMD-mediated pyroptosis. (Microbial infection) Ubiquitinated by S.flexneri IpaH7.8, leading to its degradation by the proteasome.
Function Gasdermin-D: Precursor of a pore-forming protein that plays a key role in host defense against pathogen infection and danger signals. This form constitutes the precursor of the pore-forming protein: upon cleavage, the released N-terminal moiety (Gasdermin-D, N-terminal) binds to membranes and forms pores, triggering pyroptosis. 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, CASP4 or CASP5 in response to canonical, as well as non-canonical (such as cytosolic LPS) inflammasome activators. 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. Homooligomerizes within the membrane and forms pores of 10-15 nanometers (nm) of inner diameter, allowing the release of mature interleukin-1 (IL1B and IL18) and triggering pyroptosis. Gasdermin pores also allow the release of mature caspase-7 (CASP7). In some, but not all, cells types, pyroptosis is followed by pyroptotic cell death, which is caused by downstream activation of ninjurins (NINJ1 or NINJ2), which mediate membrane rupture (cytolysis). Also forms pores in the mitochondrial membrane, resulting in release of mitochondrial DNA (mtDNA) into the cytosol. 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. Under cell culture conditions, also active against intracellular bacteria, such as Listeria monocytogenes. Also active in response to MAP3K7/TAK1 inactivation by Yersinia toxin YopJ, which triggers cleavage by CASP8 and subsequent activation. Strongly binds to bacterial and mitochondrial lipids, including cardiolipin. Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine. Gasdermin-D, p13: Transcription coactivator produced by the cleavage by CASP3 or CASP7 in the upper small intestine in response to dietary antigens. Required to maintain food tolerance in small intestine: translocates to the nucleus and acts as a coactivator for STAT1 to induce the transcription of CIITA and MHC class II molecules, which in turn induce type 1 regulatory T (Tr1) cells in upper small intestine. Gasdermin-D, p40: Produced by the cleavage by papain allergen. After cleavage, moves to the plasma membrane and homooligomerizes within the membrane and forms pores of 10-15 nanometers (nm) of inner diameter, allowing the specific release of mature interleukin-33 (IL33), promoting type 2 inflammatory immune response.
Protein Name Gasdermin-D
Gasdermin Domain-Containing Protein 1 Cleaved Into - Gasdermin-D - N-Terminal
Gsdmd-Nt
Hgsdmd-Ntd - Gasdermin-D - C-Terminal
Gsdmd-Ct
Hgsdmd-Ctd - Gasdermin-D - P13
Gasdermin-D - 13 Kda
13 Kda Gsdmd - Gasdermin-D - P40
Database Links Reactome: R-HSA-111457
Reactome: R-HSA-448706
Reactome: R-HSA-5620971
Reactome: R-HSA-5686938
Reactome: R-HSA-6798695
Reactome: R-HSA-9660826
Cellular Localisation Gasdermin-D: 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
Gasdermin-D
N-Terminal: Cell Membrane
Multi-Pass Membrane Protein
Secreted
Mitochondrion Membrane
Released In The Extracellular Milieu Following Pyroptosis
N-Terminal: Cytoplasm
(Microbial Infection) Upon Infection By M
Tuberculosis
Localization To Cell Membrane Is Prevented By M
Tuberculosis Phosphatase Ptpb That Catalyzes Dephosphorylation Of Phosphatidylinositol (4
5)-Bisphosphate And Phosphatidylinositol 4-Phosphate
Thereby Inhibiting The Membrane Targeting Of Gasdermin-D
N-Terminal And Subsequent Cytokine Release And Pyroptosis
P13: Nucleus
C-Terminal: Cytoplasm
Alternative Antibody Names Anti-Gasdermin-D antibody
Anti-Gasdermin Domain-Containing Protein 1 Cleaved Into - Gasdermin-D - N-Terminal antibody
Anti-Gsdmd-Nt antibody
Anti-Hgsdmd-Ntd - Gasdermin-D - C-Terminal antibody
Anti-Gsdmd-Ct antibody
Anti-Hgsdmd-Ctd - Gasdermin-D - P13 antibody
Anti-Gasdermin-D - 13 Kda antibody
Anti-13 Kda Gsdmd - Gasdermin-D - P40 antibody
Anti-GSDMD antibody
Anti-DFNA5L antibody
Anti-GSDMDC1 antibody
Anti-FKSG10 antibody

Information sourced from Uniprot.org

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