GSDMD Antibody | Affinity Biosciences

Product:	GSDMD Antibody
Catalog:	DF12275
Description:	Rabbit polyclonal antibody to GSDMD
Application:	WB IHC IF/ICC
Reactivity:	Human
Prediction:	Pig, Bovine, Horse, Dog
Mol.Wt.:	50kDa; 53kD(Calculated).
Uniprot:	P57764
RRID:	AB_2845080

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human

Prediction:

Pig(82%), Bovine(82%), Horse(82%), Dog(91%)

Clonality:

Polyclonal

Specificity:

GSDMD Antibody detects endogenous levels of total GSDMD.

RRID:

AB_2845080
Cite Format: Affinity Biosciences Cat# DF12275, RRID:AB_2845080.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

1810036L03Rik; DF 5L; DF5L; DFNA 5L; DFNA5L; FKSG 10; FKSG10; FLJ12150; Gasdermin D; Gasdermin domain containing 1; Gasdermin domain containing protein 1; Gasdermin domain-containing protein 1; Gasdermin-D; GasderminD; GSDMD; GSDMD_HUMAN; GSDMDC 1; GSDMDC1;

Immunogens

Immunogen:

Uniprot:

P57764(GSDMD_HUMAN) >>Visit HPA database.

Gene(ID):

GSDMD(79792)

Expression:

P57764 GSDMD_HUMAN:

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:

P57764 GSDMD_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MGSAFERVVRRVVQELDHGGEFIPVTSLQSSTGFQPYCLVVRKPSSSWFWKPRYKCVNLSIKDILEPDAAEPDVQRGRSFHFYDAMDGQIQGSVELAAPGQAKIAGGAAVSDSSSTSMNVYSLSVDPNTWQTLLHERHLRQPEHKVLQQLRSRGDNVYVVTEVLQTQKEVEVTRTHKREGSGRFSLPGATCLQGEGQGHLSQKKTVTIPSGSTLAFRVAQLVIDSDLDVLLFPDKKQRTFQPPATGHKRSTSEGAWPQLPSGLSMMRCLHNFLTDGVPAEGAFTEDFQGLRAEVETISKELELLDRELCQLLLEGLEGVLRDQLALRALEEALEQGQSLGPVEPLDGPAGAVLECLVLSSGMLVPELAIPVVYLLGALTMLSETQHKLLAEALESQTLLGPLELVGSLLEQSAPWQERSTMSLPPGLLGNSWGEGAPAWVLLDECGLELGEDTPHVCWEPQAQGRMCALYASLALLSGLSQEPH

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Dog

Pig

Horse

Bovine

Rabbit

Sheep

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P57764 As Substrate

P57764

Site	PTM Type	Source
Y37	Phosphorylation	Uniprot
K43	Ubiquitination	Uniprot
K51	Ubiquitination	Uniprot
K55	Ubiquitination	Uniprot
K62	Ubiquitination	Uniprot
S79	Phosphorylation	Uniprot
K145	Ubiquitination	Uniprot
S152	Phosphorylation	Uniprot
Y158	Phosphorylation	Uniprot
T161	Phosphorylation	Uniprot
K168	Ubiquitination	Uniprot
S181	Phosphorylation	Uniprot
S185	Phosphorylation	Uniprot
S201	Phosphorylation	Uniprot
K203	Ubiquitination	Uniprot
K204	Ubiquitination	Uniprot
K236	Ubiquitination	Uniprot
K248	Ubiquitination	Uniprot
S250	Phosphorylation	Uniprot
T251	Phosphorylation	Uniprot
S252	Phosphorylation	Uniprot
S261	Phosphorylation	Uniprot
K299	Ubiquitination	Uniprot

Research Backgrounds

P57764_GSDMD_HUMAN

Function:

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. 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, possibly allowing the release of mature IL1B and triggering pyroptosis. 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. 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.

PTMs:

Cleavage at Asp-275 by CASP1 (mature and uncleaved precursor forms) or CASP4 relieves autoinhibition and is sufficient to initiate pyroptosis. Cleavage at Asp-87 by CASP3.

Subcellular Location:

Cytoplasm>Cytosol. Inflammasome.
Note: In response to a canonical inflammasome stimulus, such as nigericin, recruited to NLRP3 inflammasone with similar kinetics to that of uncleaved CASP1 precursor.

Cell membrane. Secreted.
Note: Released in the extracellular milieu following pyroptosis.

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.

Subunit Structure:

In response to a canonical inflammasome stimulus, such as nigericin, recruited to NLRP3 inflammasone with similar kinetics to that of uncleaved CASP1 precursor. Although this recruitment is also observed in the absence of PYCARD, it is more efficient in its presence (By similarity). Gasdermin-D, N-terminal forms disulfide-linked homooligomers (16-mers) in a Ca(+2)-independent manner. Oligomerization occurs in the presence of membranes; cytosolic Gasdermin-D, N-terminal remains monomeric.

Family&Domains:

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.

Belongs to the gasdermin family.

Research Fields

· Organismal Systems > Immune system > NOD-like receptor signaling pathway. (View pathway)

References

1). Elamipretide alleviates pyroptosis in traumatically injured spinal cord by inhibiting cPLA2-induced lysosomal membrane permeabilization. Journal of Neuroinflammation (PubMed: 36609266) [IF=9.3]

2). Shionone alleviates NLRP3 inflammasome mediated pyroptosis in interstitial cystitis injury. International Immunopharmacology (PubMed: 33223465) [IF=5.6]

3). Melatonin protects human nucleus pulposus cells from pyroptosis by regulating Nrf2 via melatonin membrane receptors. Bone & Joint Research (PubMed: 37051810) [IF=4.6]

4). Staphylococcus aureus mediates pyroptosis in bovine mammary epithelial cell via activation of NLRP3 inflammasome. Veterinary Research (PubMed: 35123552) [IF=4.4]

Application: WB Species: bovine Sample: MAC-T cells

Figure 1 GSDMD activation by S. aureus. A MAC-T cells were treated with S. aureus for the indicated times. GSDMD and GSDMD-N were detected by immunoblotting. B Cell death was measured by LDH release. C Subcellular localization of GSDMD-N or GSDMD in MAC-T cells incubated with or without S. aureus.

Application: IF/ICC Species: bovine Sample: MAC-T cells

5). Dihydromyricetin inhibits African swine fever virus replication by downregulating toll-like receptor 4-dependent pyroptosis in vitro. Veterinary Research (PubMed: 37438783) [IF=4.4]

Application: WB Species: pig Sample:

Figure 7 DHM treatment inhibited ASFV-induced pyroptosis. PAMs were treated with A and B DHM or C disulfiram after ASFV infection; the cells were then collected at 24 h for Western blotting or RT-qPCR. D and E Western blotting of ASFV-infected PAMs were treated with DHM in the presence or absence of D polyphyllin VI or E RS 09 TFA; samples were collected after 24 h of treatment. *P

6). Tryptophan alleviates lipopolysaccharide-induced liver injury and inflammation by modulating necroptosis and pyroptosis signaling pathways in piglets. Animal Biotechnology (PubMed: 37688392) [IF=3.7]

Application: WB Species: piglets Sample: liver

Figure 7. Effects of tryptophan on expression pyroptosis-related proteins in liver after 4h lipopolysaccharide (LPS) challenge in piglets. P values < 0.05 were considered significant. P values between 0.05 and 0.10 were regarded as a tendency. Values are presented as mean ± standard error, n = 4. CONTR: control group; LPS: piglets challenged with LPS; Trp + LPS: piglets fed with 0.2% tryptophan and challenged with LPS. NLRP3: NOD-like receptor family pyrin domain containing 3. GSDMD: gasdermin-D. MLKL: the mixed lineage kinase domain-like protein. HSP70: the 70-kDa heat shock proteins.

7). Oxalate‑induced renal pyroptotic injury and crystal formation mediated by NLRP3‑GSDMD signaling in vitro and in vivo. Molecular Medicine Reports (PubMed: 37732544) [IF=3.4]

Application: IHC Species: Human Sample: HK-2 cells

Figure 7. GSDMD mediates oxalate crystal-related pyroptosis in vivo. (A) Von Kossa staining revealed oxalate crystals in kidney of mice treated with Gly. (B) Haematoxylin and eosin staining demonstrated renal epithelial cell swelling, renal tubule edema and neutrophil infiltration in Gly treated mice. Scale bars, 50 µm. (C) Serum creatinine level of Gly-treated mice was examined by creatinine assay (n=6). (D) Western blot analysis of indicated proteins in NLRP3-GSDMD pathway in oxalated crystal mice. (E) The expression of same proteins was detected through immunohistochemical assays in mice. Scale bars, 50 µm. GSDMD, gasdermin D; Gly, glyoxylic acid; NLRP3, leucine-rich repeat-containing family pyrin domain-containing 3; OPN, osteopontin; Ctrl, control.

Application: WB Species: Human Sample: HK-2 cells

Figure 3. NLRP3 knockdown ameliorates oxalate-caused injury of HK-2 cells. (A) Western blot detection of NLRP3, caspase-1, GSDMD and GSDMD-N protein contents in HK-2 cells treated with 0.8 mM oxalate for 24 h after transfection with NLRP3-siRNA. (B) Relative lactate dehydrogenase levels in HK-2 cells transfected with NLRP3-siRNA upon 0.8 mM oxalate treatment for 24 h (n=6). (C) Images of two colors (blue and green) and merged colors in HK-2 cells with TUNEL staining were observed by confocal microscopy. HK-2 cells with NLRP3 silence were treated with 0.8 mM oxalate for 24 h followed by staining. Scale bars, 100 µm. (D) Images of three colors (blue, green and red) and merged colors in HK-2 cells with Calcein-AM/PI staining were recorded through confocal microscopy. HK-2 cells transfected with NLRP3-siRNA or scrambled-siRNA were dealt with 0.8 mM oxalate for 24 h. Scale bars, 100 µm. NLRP3, leucine-rich repeat-containing family pyrin domain-containing 3; GSDMD, gasdermin D; siRNA, small interfering RNA; Ctrl, control; NC, negative control.

8). LPS Enhances the Chemosensitivity of Oxaliplatin in HT29 Cells via GSDMD-Mediated Pyroptosis. Cancer Management and Research (PubMed: 33116894) [IF=3.3]

Application: WB Species: mouse Sample: colonials

Figure 1 |LPS improves anti-cancer effect of oxaliplatin in mice in vivo experiment.(D–F) Histopathology of colonials in different phases of model establishment. Western blot analyses and IHC of GSDMD and GSDMD-N in each group (n=6).

Application: IHC Species: human Sample: colorectal cancer tissues

Figure 2| The GSDMD expression is reduced in human colorectal cancer tissues and is significantly related to the overall survival of CRC patients.(C) Immunohistochemical analysis of GSDMD expression in human colorectal cancer tissues (n=244).

9). Immunohistochemical analysis of pyroptosis‐related protein expression in IgG4‐related sialadenitis. Journal of Oral Pathology & Medicine (PubMed: 35218237) [IF=3.3]

Application: IHC Species: Human Sample: IgG4‐RS tissues

FIGURE 1 Immunohistochemical (IHC) staining of pyroptosis‐related proteins in IgG4‐related sialadenitis (IgG4‐RS) tissues and control tissues (400×). IHC staining of IgG4‐RS tissues: (A) NLRP3 (NOD‐, LRR‐ and pyrin domain‐containing protein); (B) ASC (apoptosis‐associated speck‐like protein containing a CARD); (C) Caspase‐1; (D) GSDMD (gasdermin family members, including digestive dermatin D; (E) interleukin 1β (IL‐1β); (F) interleukin 18 (IL‐18). IHC staining of the control tissues: (a) NLRP3; (b) ASC; (c) Caspase‐1; (d) GSDMD; (e) IL‐1β; (f) IL‐18. Scale bars, 100 μm

10). Transcription factor Krüppel-like factor 4 upregulated G protein-coupled receptor 30 alleviates intestinal inflammation and apoptosis, and protects intestinal integrity from intestinal ischemia–reperfusion injury. Immunity, Inflammation and Disease (PubMed: 37506161) [IF=3.2]

Application: WB Species: Human Sample: Caco‐2 cells.

Figure 7 KLF4/GPR30 regulates NLRP3 inflammasome and pyroptosis in OGD/R‐exposed Caco‐2 cells. (A) Untransfected and transfected Caco‐2 cells were exposed to OGD/R, and protein expression of NLRP3, cleaved‐caspase1, caspase1, and GSDMD‐N was measured using western blot. (B) Immunofluorescence was conducted to examine GSDMD‐N expression. OGD/R, oxygen‐glucose deprivation/reoxygenation. ***p

Application: IF/ICC Species: Human Sample: Caco‐2 cells.

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For Research Use Only.
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GSDMD Antibody - #DF12275