GSDMD Antibody | Affinity Biosciences

Product:	GSDMD Antibody
Catalog:	AF4012
Description:	Rabbit polyclonal antibody to GSDMD
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Horse
Mol.Wt.:	53 kDa(full), 35kDa(cleaved); 53kD(Calculated).
Uniprot:	P57764
RRID:	AB_2846776

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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

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 1:50-1:200
*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,Mouse,Rat

Prediction:

Pig(88%), Horse(83%)

Clonality:

Polyclonal

Specificity:

GSDMD Antibody detects endogenous levels of total GSDMD protein, including precursor and the cleaved N-terminal fragment.

RRID:

AB_2846776
Cite Format: Affinity Biosciences Cat# AF4012, RRID:AB_2846776.

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

Pig

Horse

Bovine

Sheep

Dog

Rabbit

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). Single-Cell Atlas of Human Ovaries Reveals The Role Of The Pyroptotic Macrophage in Ovarian Aging. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2024 (PubMed: 38036420) [IF=15.1]

Application: IHC Species: Human Sample:

Figure 5 Pyroptotic macrophages promote stromal cell senescence. A) Representative images of senescence‐associated β‐galactosidase (SA‐beta‐gal) staining of stromal cells treated with the supernatant of TNF‐α/IFN‐γ‐stimulated THP1 cells, the supernatant of LPS/Nig‐stimulated THP1 cells or the supernatant of unstimulated THP1 cells (scale bar, 500 µm). Statistical analyses of SA‐beta‐gal‐positive cell rates were shown in the right image. B) The relative expression of fibrosis‐related genes in ovarian stromal cells treated with supernatant of unstimulated or stimulated THP1 cells. The results were analyzed by ANOVA. C) Representative IHC staining for GSDMD showing the elevated expression of GSDMD in the middle‐aged group (scale bar, 100 µm). The insert at top left corner of each image is the magnified image (scale bar, 250 µm). D) EREG concentration in follicular fluids measured by ELISA. E) Flow cytometric analysis of follicular fluids from IVF patients: representative flow cytometric dot plots of macrophages in follicular fluids from each group. Statistical graph of the percentages of macrophages in follicular fluids. F) Representative histogram plots and the statistical graph of the MFI of GSDMD. MFI: mean fluorescence intensity.

Application: IF/ICC Species: Human Sample:

Figure 6 The inhibition of pyroptosis impedes ovarian aging and promotes female health. A) The expression levels of pyroptosis‐related proteins in ovaries from naturally aged (old), prematurely aged (POI) and LPS‐stimulated mice were detected by western blotting. B) Representative immunofluorescence images of GSDMD staining of ovarian sections from young and old mice, with nuclei counterstained with DAPI (scale bar, 50 µm). C) Representative HE‐stained ovarian tissue sections from mice of different genotypes at the ages of 3 months (3 mo), 8 months (8 mo), and 13 months (13 mo). D) The total number of follicles of each stage was counted and statistically analyzed. The error bars indicate SD. *p

2). Adipocyte pyroptosis occurs in omental tumor microenvironment and is associated with chemoresistance of ovarian cancer. Journal of biomedical science, 2024 (PubMed: 38862973) [IF=11.0]

3). Hair follicle-MSC-derived small extracellular vesicles as a novel remedy for acute pancreatitis. JOURNAL OF CONTROLLED RELEASE, 2022 (PubMed: 36402231) [IF=10.8]

4). Betulinic Acid Inhibits ROS-Mediated Pyroptosis in Spinal Cord Injury by Augmenting Autophagy via the AMPK-mTOR-TFEB Signaling Pathway. International Journal of Biological Sciences, 2020 (PubMed: 33867836) [IF=9.2]

Application: WB Species: Mice Sample: spinal cords

Figure 4 Inhibition of autophagy reverses the effects of BA on pyroptosis after SCI. (A) Immunofluorescence staining for p62 and NeuN co-localization at the spinal cord lesion after SCI (scale bar = 25 µm). (B) The quantitative mean optical density of the p62 in motor neurons of spinal cord lesion in each group. (C) Immunofluorescence staining for LC3II and NeuN co-localization at the spinal cord lesion after SCI (scale bar = 25 µm). (D) The quantitative mean number of the LC3II positive neurons in motor neurons of spinal cord lesion in each group. (E) Western blotting for the p62, LC3II, Beclin1, Vps34, and CTSD expression levels in the BA and BA+3MA groups. The gels were run under the same experimental conditions, and the cropped blots are shown here. (F) The optical density values of the p62, LC3II, Beclin1, Vps34, and CTSD expression levels were quantified and analyzed in each group. (G) Immunofluorescence staining for Caspase-1 and NeuN co-localization in the spinal cords of the BA and BA+3MA groups (scale bar = 25 µm) (H) The quantitative mean optical density of the Caspase-1 in motor neurons of spinal cord lesion. (I) Immunofluorescence staining for GSDMD and NeuN co-localization in the spinal cords of the BA and BA+3MA groups (scale bar = 25 µm) (J) The quantitative mean optical density of the GSDMD in motor neurons of spinal cord lesion. (K) Western blotting for the ASC, Caspase-1, GSDMD, IL-1β, IL-18 and NLRP3 expression levels in the BA and BA+3MA groups. The gels were run under the same experimental conditions, and the cropped blots are shown here. (L) The optical density values of the ASC, Caspase-1, GSDMD, IL-1β, IL-18 and NLRP3 expression levels were quantified and analyzed in each group. The values are expressed as the means ± SEM, n=5 per group. *p< 0.05 and **p< 0.01, vs. BA group.

5). Complement induces podocyte pyroptosis in membranous nephropathy by mediating mitochondrial dysfunction. Cell Death & Disease, 2022 (PubMed: 35351877) [IF=9.0]

6). Acetyl-CoA synthetase 2 induces pyroptosis and inflammation of renal epithelial tubular cells in sepsis-induced acute kidney injury by upregulating the KLF5/NF-κB pathway. Cell communication and signaling : CCS, 2024 (PubMed: 38515158) [IF=8.4]

Application: WB Species: Mouse Sample: kidney

Fig. 1 The NLRP3 inflammasome and pyroptosis are activated in an LPS-induced mouse sepsis model. A Serum creatinine levels in vehicle-treated mice and LPS-induced sepsis mice were measured (n = 5-6). B H&E staining of kidney tissues from the AKI model mice and control mice (Scale bars, 50 μm). C F4/80 staining of kidney tissues of the noted mice (Scale bars, 200 μm). D Expression of pro-inflammatory factors at the mRNA level in the kidneys of vehicle- or LPS-treated mice were determined using RT-qPCR (n = 5). E The protein expression of NLRP3, cleaved GSDMD, and caspase-1 in the kidney were detected by immunofluorescent staining (green, caspase-1; red, NLRP3 or cleaved GSDMD; blue, DAPI) and Western blotting (n = 3). Data were presented as mean ± SD. *P

7). WTAP-mediated N6-methyladenosine modification of NLRP3 mRNA in kidney injury of diabetic nephropathy. CELLULAR & MOLECULAR BIOLOGY LETTERS, 2022 (PubMed: 35761192) [IF=8.3]

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

"Fig. 2 Knockdown of WTAP inhibits high glucose (HG)-induced cell pyroptosis and pro-inflammatory factor release in HK-2 cells. A Cell viability; (B, C) cell pyroptosis; (D) LDH activity; E, F expression of WTAP, NLRP3, pro-caspase-1, active caspase-1, GSDMD, and GSDMD-N; and (G, H) the release of IL-18, IL-1β, TNF-α, and IL-6 in HK-2 cells treated with NG or HG with or without WTAP knockdown (n = 3). One-way analysis of variance was used to analyze the data among multiple groups, followed by Tukey’s post hoc test. ***P < 0.001 compared with NG + shNC. ##P < 0.01, ###P < 0.001 compared with HG + shNC"

8). A congenital CMV infection model for follow-up studies of neurodevelopmental disorders, neuroimaging abnormalities, and treatment. JCI Insight, 2022 (PubMed: 35014624) [IF=8.0]

Application: WB Species: Mice Sample: brain

Figure 9 Impact of neonatal GCV treatment on viral load, inflammation, and pyroptosis. (A) Timeline of GCV treatment. GCV was injected i.p. daily from P1 to P7. Experimental groups included (i) GCV-treated mock-infected (Mock), (ii) PBS-treated MCMV-infected (MCMV, negative treatment control), and (iii) GCV-treated MCMV-infected (MCMV+GCV) mice. (B) GCV treatment and viral loads in brain. Viral genome copy number was assessed by qPCR using DNA extracted from equal amounts of cerebral samples at the indicated time points. Data are from 3 mice/group in 3 independent experiments and analyzed by Student’s t test. (C) GCV treatment and cytokine levels. Levels of IL-1β, IL-18, TNF-α, and IFN-γ in cerebral samples were determined from 6 newborns/group in 3 independent experiments and analyzed by Student’s t test. (D) GCV treatment and immune cells infiltration. Percentages of leukocytes; B, T, and NK cells; neutrophils; monocytes; microglia; and macrophages in total cells of the cerebrum from P7 newborns were assessed by flow cytometry. Data were collected from 12 newborns/group and analyzed by Student’s t test. (E) GCV treatment and pyroptosis. Both full-length and active GSDMD (GSDMD-FL and -N) and IE1 (indicated by arrows) in cerebral lysates were detected by Western blot. GAPDH served as a loading control. For all statistical tests, results are presented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

9). Glyburide confers neuroprotection against age-related macular degeneration (AMD). Translational research : the journal of laboratory and clinical medicine, 2024 (PubMed: 38815899) [IF=7.8]

10). Inhibition of the NLRP3/caspase-1 signaling cascades ameliorates ketamine-induced renal injury and pyroptosis in neonatal rats. BIOMEDICINE & PHARMACOTHERAPY, 2022 (PubMed: 35679721) [IF=7.5]

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GSDMD Antibody - #AF4012