Gasdermin D Recombinant Rabbit Monoclonal Antibody [PD00-18] - BSA and Azide free
Catalog# HA750486
Gasdermin D Recombinant Rabbit Monoclonal Antibody [PD00-18] - BSA and Azide free
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WB
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IHC-P
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IF-Cell
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IF-Tissue
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Gasdermin D Recombinant Rabbit Monoclonal Antibody [PD00-18] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Gasdermin D full length protein.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, IF-Tissue
Target Molecular Weight
Predicted band size: 53/30 kDa
Positive Control
SiHa cell lysate, Jurkat cell lysate, THP-1 cell lysate, PC-3M cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, L6 cell lysate, THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate, mouse intestine tissue, L6.
Conjugation
unconjugated
Clone Number
PD00-18
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
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 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.
Background References
1. Sborgi L. et. al. GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J. 35:1766-1778(2016).
2. Ding J. et. al. Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535:111-116(2016).
Subcellular Location
Cell membrane, Secreted.
Synonyms
1810036L03Rik antibody
DF 5L antibody
DF5L antibody
DFNA 5L antibody
DFNA5L antibody
FKSG 10 antibody
FKSG10 antibody
FLJ12150 antibody
Gasdermin antibody
Gasdermin D antibody
ExpandImages
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This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA721144) at 1/2,000 dilution.
Lane 1: SiHa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: THP-1 cell lysate
Lane 4: PC-3M cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: PC-12 cell lysate
Lane 7: L6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 53/30 kDa
Observed band size: 53/30 kDa
Exposure time: 43 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721144) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA721144) at 1/2,000 dilution.
Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate (20 µg/Lane)
Predicted band size: 53/30 kDa
Observed band size: 53/35 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721144) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse intestine tissue with Rabbit anti-Gasdermin D antibody (HA721144) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721144) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of L6 cells labeling Gasdermin D with Rabbit anti-Gasdermin D antibody (HA721144) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Gasdermin D antibody (HA721144) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Intestine
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA721144, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
☑ Knockout (KO),Cell treatment (CT)
This data was developed using HA721144, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA721144) at 1/2,000 dilution.
Lane 1: HCT-8 WT
Lane 2: HCT-8 GSDMD KO
Lane 3: HCT-8 GSDMD KO+empty vector
Lane 4: HCT-8 GSDMD KO+OE-GSDMD
Lysates/proteins at 20 µg/Lane.
Exposure time: 59 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721144, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 53 kDa
Observed band size: 53 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Alternative Products
Cleaved Gasdermin D (Asp275) Recombinant Rabbit Monoclonal Antibody [PSH10-87]
Application: WB,IP
Reactivity: Human
Conjugate: unconjugated
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