GSDMD Antibody - #AF4012

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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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 100ul $280 In stock
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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:
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
88
Horse
83
Bovine
75
Sheep
75
Dog
71
Rabbit
71
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P57764 As Substrate

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

Research Backgrounds

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.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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). Hair follicle-MSC-derived small extracellular vesicles as a novel remedy for acute pancreatitis. JOURNAL OF CONTROLLED RELEASE (PubMed: 36402231) [IF=10.8]
2). 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 (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.
3). Complement induces podocyte pyroptosis in membranous nephropathy by mediating mitochondrial dysfunction. Cell Death & Disease (PubMed: 35351877) [IF=9.0]
4). WTAP-mediated N6-methyladenosine modification of NLRP3 mRNA in kidney injury of diabetic nephropathy. CELLULAR & MOLECULAR BIOLOGY LETTERS (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"
5). A congenital CMV infection model for follow-up studies of neurodevelopmental disorders, neuroimaging abnormalities, and treatment. JCI Insight (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.
6). Inhibition of the NLRP3/caspase-1 signaling cascades ameliorates ketamine-induced renal injury and pyroptosis in neonatal rats. BIOMEDICINE & PHARMACOTHERAPY (PubMed: 35679721) [IF=7.5]
7). Exercise induced meteorin-like protects chondrocytes against inflammation and pyroptosis in osteoarthritis by inhibiting PI3K/Akt/NF-κB and NLRP3/caspase-1/GSDMD signaling. Biomedicine & Pharmacotherapy (PubMed: 36527845) [IF=7.5]
8). Ezrin inhibition alleviates oxidative stress and pyroptosis via regulating TRPML1-calcineurin axis mediated enhancement of autophagy in spinal cord injury. Free radical biology & medicine (PubMed: 38142951) [IF=7.4]
9). Wedelolactone alleviates acute pancreatitis and associated lung injury via GPX4 mediated suppression of pyroptosis and ferroptosis. Free Radical Biology and Medicine (PubMed: 34246777) [IF=7.4]

Application: WB    Species: Rat    Sample: AR42J cells

Fig. 5. Wed suppressed pancreatic pyroptosis. (a) The expression of pyroptosis-associated proteins was detected by Western blot. (b, c) The serum IL-1β and IL-18 levels in Taru-induced AP rats. (d, e) The serum IL-1β and IL-18 levels in Caer-induced AP rats. **P < 0.01 vs. Sham group, §§ P < 0.01 vs. Taur group, †† P < 0.01 vs. Control group, && P < 0.01 vs. Caer group.
10). Dietary disodium fumarate supplementation alleviates subacute ruminal acidosis (SARA)-induced liver damage by inhibiting pyroptosis via mitophagy-NLRP3 inflammasome pathway in lactating Hu sheep. Frontiers in Immunology (PubMed: 37275885) [IF=7.3]

Application: WB    Species: Hu Sheep    Sample: liver tissue

Figure 4 Effect of DF supplementation on SARA-induced pyroptosis in the liver of Hu sheep. The expression of pyroptosis-related (A) gene and (B, C) protein in the liver tissue of Hu sheep were detected. n=6 per group and the values are the mean ± SEM. ** P
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