概述
产品名称
Gasdermin D (N terminal) Rabbit Polyclonal Antibody
抗体类型
Rabbit Polyclonal Antibody
免疫原
Recombinant protein within human Gasdermin D aa 100-300.
种属反应性
Human, Mouse, Rat
验证应用
WB, IHC-P, FC, IF-Cell
分子量
Predicted band size: 53 kDa
阳性对照
SiHa cell lysate, PC-3 cell lysate, Jurkat cell lysate, THP-1 cell lysate, human kidney tissue lysate, SiHa, rat stomach tissue, human tonsil tissue, human prostate carcinoma tissue, human esophagus tissue, mouse colon tissue, mouse intestine tissue, SiHa, PC-3M, RAW264.7, L6.
偶联
unconjugated
RRID
产品特性
形态
Liquid
浓度
1ug/ul
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Immunogen affinity purified.
应用稀释度
-
WB
-
1:500-1:2,000
-
IHC-P
-
1:50-1:1,000
-
FC
-
1:1,000
-
IF-Cell
-
1:100
发表文章中的应用
发表文章中的种属
靶点
功能
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.
背景文献
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).
序列相似性
Belongs to the gasdermin family.
组织特异性
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.
翻译后修饰
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.
亚细胞定位
Cytosol. Secreted. Plasma membrane.
别名
1810036L03Rik antibody
DF 5L antibody
DF5L antibody
DFNA 5L antibody
DFNA5L antibody
FKSG 10 antibody
FKSG10 antibody
FLJ12150 antibody
Gasdermin antibody
Gasdermin D antibody
展开1810036L03Rik antibody
DF 5L antibody
DF5L antibody
DFNA 5L antibody
DFNA5L antibody
FKSG 10 antibody
FKSG10 antibody
FLJ12150 antibody
Gasdermin antibody
Gasdermin D antibody
Gasdermin domain containing 1 antibody
Gasdermin domain containing protein 1 antibody
Gasdermin domain-containing protein 1 antibody
Gasdermin-D antibody
GasderminD antibody
GSDMD antibody
GSDMD_HUMAN antibody
GSDMDC 1 antibody
GSDMDC1 antibody
折叠图片
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Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) at 1/1,000 dilution.
Lane 1: SiHa cell lysate (10 µg/Lane)
Lane 2: PC-3 cell lysate (10 µg/Lane)
Lane 3: Jurkat cell lysate (10 µg/Lane)
Lane 4: THP-1 cell lysate (10 µg/Lane)
Lane 5: Human kidney tissue lysate (20 µg/Lane)
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 2 minutes;
12% 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 (ER1901-37) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Gasdermin D (N terminal) on THP-1 cell lysates with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) at 1/1,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 53 kDa
Observed band size: 53/35 kDa
Exposure time: 25 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 (ER1901-37) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded rat stomach tissue using anti-Gasdermin D (N terminal) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-37, 1/200) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Gasdermin D (N terminal) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-37, 1/50) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-Gasdermin D (N terminal) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-37, 1/50) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Gasdermin D (N terminal) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-37, 1/200) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using anti-Gasdermin D (N terminal) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-37, 1/200) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded mouse intestine tissue with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) 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 (ER1901-37) 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. -
Immunocytochemistry analysis of PC-3M cells labeling Gasdermin D (N terminal) with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (N terminal) antibody (ER1901-37) 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 (M1305-2, 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. -
Immunocytochemistry analysis of RAW264.7 cells labeling Gasdermin D (N terminal) with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (N terminal) antibody (ER1901-37) 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 (M1305-2, 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. -
Immunocytochemistry analysis of L6 cells labeling Gasdermin D (N terminal) with Rabbit anti-Gasdermin D (N terminal) antibody (ER1901-37) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (N terminal) antibody (ER1901-37) 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 (M1305-2, 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. -
Flow cytometric analysis of Gasdermin D (N terminal) was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-37, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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引文
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