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AB324385

Anti-GSDMD antibody [EPR30356-588]

  • BOND RX™ Validated
  • RabMAb
  • Recombinant
  • 20ul selling size
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Rabbit Recombinant Monoclonal GSDMD antibody. Suitable for IHC-P, WB and reacts with Human samples.

View Alternative Names

DFNA5L, GSDMDC1, FKSG10, GSDMD, Gasdermin-D, Gasdermin domain-containing protein 1

7 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Immunohistochemical analysis of paraffin-embedded (A) Adjacent human liver and (B) Human hepatocellular carcinoma tissue labeling GSDMD with ab324385 at 1/1000 (0.522 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in (A) adjacent human liver, with increased expression of GSDMD in (B) hepatocellular carcinoma tissue.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Immunohistochemical analysis of paraffin-embedded (A) Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cell pellet, (B) PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet, (C) IMR-32 (human neuroblastoma neuroblast) cell pellet and (D) HEK-293T (human embryonic kidney epithelial cell) cell pellet labeling GSDMD with ab324385 at 1/1000 (0.522 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on (A) Jurkat and (B) PC-3 cell pellet, no staining on (C) IMR-32 and (D) HEK-293T cell pellet.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling GSDMD with ab324385 at 1/1000 (0.522 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human cardiac muscle.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Immunohistochemical analysis of paraffin-embedded (A) Adjacent human colon and (B) Human colon carcinoma tissue labeling GSDMD with ab324385 at 1/1000 (0.522 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in (A) adjacent human colon, with increased expression of GSDMD in (B) colon carcinoma tissue.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • WB

Supplier Data

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

GSDMD is cleaved in LPS treated cells (PMID : 26375003).

The lysates were kindly provided by Dr Feng Shao's lab NIBS

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GSDMD antibody [EPR30356-588] (ab324385) at 1/1000 dilution

Lane 1:

Untreated AML2 (human acute myeloid leukemia cell) whole cell lysate at 20 µg

Lane 2:

AML2 treated with 5µg/ml LPS and 5µg/ml nigericin for 1 hour whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 52 kDa,23 kDa,36 kDa

false

Exposure time: 1s

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • WB

Supplier Data

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

GSDMD is cleaved in LPS treated cells (PMID : 26375003).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GSDMD antibody [EPR30356-588] (ab324385) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

THP-1 treated with 50ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1µg/ml BFA for another 3 h whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 52 kDa,23 kDa,37 kDa

false

Exposure time: 15s

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)
  • WB

Supplier Data

Western blot - Anti-GSDMD antibody [EPR30356-588] (AB324385)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : IMR-32, 293T

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-4 : 6 seconds, lane 5-6 : 59 seconds

All lanes:

Western blot - Anti-GSDMD antibody [EPR30356-588] (ab324385) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 52 kDa,37 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR30356-588

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

WB, IHC-P

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

This antibody can detect full length, as well as a C-terminal fragment after stimulation in WB.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/1000", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" } } }
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Product details

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Gasdermin-D. Precursor of a pore-forming protein that plays a key role in host defense against pathogen infection and danger signals (PubMed : 26375003, PubMed : 26375259, PubMed : 27281216). 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 (PubMed : 26375003, PubMed : 26375259, PubMed : 27281216).. Gasdermin-D, N-terminal. Promotes pyroptosis in response to microbial infection and danger signals (PubMed : 26375003, PubMed : 26375259, PubMed : 27418190, PubMed : 28392147, PubMed : 32820063, PubMed : 34289345, PubMed : 38040708, PubMed : 38530158, PubMed : 38599239). 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 (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, PubMed : 29898893, PubMed : 36227980). 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 (PubMed : 27281216, PubMed : 27418190, PubMed : 29898893, PubMed : 33883744, PubMed : 38040708, PubMed : 38530158, PubMed : 38599239). Gasdermin pores also allow the release of mature caspase-7 (CASP7) (By similarity). In some, but not all, cells types, pyroptosis is followed by pyroptotic cell death, which is caused by downstream activation of ninjurin-1 (NINJ1), which mediates membrane rupture (cytolysis) (PubMed : 33472215, PubMed : 37198476). Also forms pores in the mitochondrial membrane, resulting in release of mitochondrial DNA (mtDNA) into the cytosol (By similarity). 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). Also active in response to MAP3K7/TAK1 inactivation by Yersinia toxin YopJ, which triggers cleavage by CASP8 and subsequent activation (By similarity). Required for mucosal tissue defense against enteric pathogens (By similarity). Activation of the non-canonical inflammasome in brain endothelial cells can lead to excessive pyroptosis, leading to blood-brain barrier breakdown (By similarity). Strongly binds to bacterial and mitochondrial lipids, including cardiolipin (PubMed : 27281216). Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine (PubMed : 27281216).. Gasdermin-D, p13. Transcription coactivator produced by the cleavage by CASP3 or CASP7 in the upper small intestine in response to dietary antigens (By similarity). 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 (By similarity).. Gasdermin-D, p40. Produced by the cleavage by papain allergen (PubMed : 35794369). 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 (PubMed : 35794369).
See full target information GSDMD
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Product promise

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