GSDMD (N-terminal) 抗体 (YA5502) (HY-P85810)

Gsdmdc1, Gsdmd, Gasdermin-D, Gasdermin domain-containing protein 1

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反应种属：Human, Mouse, Rat

抗原名称：GSDMD

应用：WB, ELISA, IF/ICC

标记：Unconjugated

用户验证的应用详细信息背景信息

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经验证应用

项目	详情
WB	Western blot analysis of extracts from PC-3 (lane 2(20μg), NIH/3T3 (lane 3(20μg), THP-1 (lane 4(20μg), Jurkat (lane 5(20μg) using GSDMD Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature. Western blot analysis of extracts from THP-1 (lane 2, 20ug), THP-1 treated with 25ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1ug/ml BFA for another 3 hours (lane3, 20μg) and THP-1 treated with 50ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1ug/ml BFA for another 3 hours (lane4, 20μg) using GSDMD (N-terminal) Antibody ( HY-P85810 ). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (HY-P8004,1/10,000) was used for 1 hour at room temperature.
ICC	Immunocytochemistry analysis of Hela cells labeling GSDMD (N-terminal) with GSDMD (N-terminal) Antibody ( HY-P85810 ) 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/100 dilution in quick block buffer overnight at 4 ℃.AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005, Green) 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. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of Hela cells labeling GSDMD (N-terminal) with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/300 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/300 dilution in quick block buffer overnight at 4 ℃.AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005, Green) 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. The Nuclear counterstain was DAPI (Blue).

项目

详情

Western blot analysis of extracts from PC-3 (lane 2(20μg), NIH/3T3 (lane 3(20μg), THP-1 (lane 4(20μg), Jurkat (lane 5(20μg) using GSDMD Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature. Western blot analysis of extracts from THP-1 (lane 2, 20ug), THP-1 treated with 25ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1ug/ml BFA for another 3 hours (lane3, 20μg) and THP-1 treated with 50ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1ug/ml BFA for another 3 hours (lane4, 20μg) using GSDMD (N-terminal) Antibody ( HY-P85810 ). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (HY-P8004,1/10,000) was used for 1 hour at room temperature.

ICC

Immunocytochemistry analysis of Hela cells labeling GSDMD (N-terminal) with GSDMD (N-terminal) Antibody ( HY-P85810 ) 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/100 dilution in quick block buffer overnight at 4 ℃.AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005, Green) 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. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of Hela cells labeling GSDMD (N-terminal) with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/300 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with GSDMD (N-terminal) Antibody ( HY-P85810 ) at 1/300 dilution in quick block buffer overnight at 4 ℃.AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005, Green) 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. The Nuclear counterstain was DAPI (Blue).

详细信息

免疫原AA range: 1-100

宿主/亚型Mouse / IgG

抗体类型Monoclonal

产品形式溶液

偶联物Non-conjugated

纯化方式affinity chromatography.

浓度不同批次浓度可能有所不同，具体浓度请参见对应批次的COA。批次浓度查询

储存缓冲液Supplied in PBS, 50% glycerol, 0.05% Proclin 300, 0.05%BSA

储存条件保存条件 & 期限: Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles. 运输条件: Shipping with blue ice.

RRIDAB_3719050

UniProt IDP57764

Gene ID79792 [ NCBI ]

背景信息

功能： GSDMD 是一种成孔蛋白前体，在宿主抵御病原体感染和危险信号的过程中发挥关键作用。该蛋白是成孔蛋白的前体：经切割后，释放的 N 端结构域 (Gasdermin-D N 端) 会结合细胞膜并形成孔道，引发焦亡；它可响应微生物感染和危险信号，促进焦亡发生。当细胞响应经典以及非经典 (如胞质 LPS) 炎症小体激活剂时，炎性半胱天冬酶 CASP1、CASP4 或 CASP5 会切割 Gasdermin-D，生成该活性结构域。切割后，该结构域会转移到质膜，与膜内侧的脂质强结合，包括单磷酸化磷脂酰肌醇 (如磷脂酰肌醇 4-磷酸)、双磷酸化磷脂酰肌醇 (如磷脂酰肌醇 (4,5)-二磷酸、磷脂酰肌醇 (3,4,5)-三磷酸)，与磷脂酸和磷脂酰丝氨酸的结合力较弱。它会在膜内同源寡聚化，形成内径 10-15 纳米 (nm) 的孔道，使成熟的白细胞介素-1 (IL1B 和 IL18) 释放，触发焦亡。Gasdermin 孔道还可使成熟的半胱天冬酶-7 (CASP7) 释放 (依据同源性推测)。在部分 (而非全部) 细胞类型中，焦亡发生后会出现焦亡性细胞死亡，这是由下游 ninjurin-1 (NINJ1) 激活介导的，NINJ1 可介导膜破裂 (细胞溶解)。它还会在线粒体外膜形成孔道，导致线粒体 DNA (mtDNA) 释放到胞质中 (依据同源性推测)。从焦亡细胞释放到胞外环境的 Gasdermin-D N 端可快速结合并杀死革兰氏阴性菌和革兰氏阳性菌，且不会损伤邻近的哺乳动物细胞，因为其脂质结合特异性使其不会从外侧破坏质膜。在细胞培养条件下，它对李斯特菌等胞内菌也有活性 (依据同源性推测)。它还可响应耶尔森菌毒素 YopJ 对 MAP3K7/TAK1 的灭活作用，此时 CASP8 会切割 GSDMD 使其激活 (依据同源性推测)。它是黏膜组织抵御肠道病原体的必要组分 (依据同源性推测)。脑内皮细胞中非经典炎症小体的激活会导致过度焦亡，造成血脑屏障破坏 (依据同源性推测)。它可与细菌和线粒体脂质 (包括心磷脂) 强结合，不结合未磷酸化的磷脂酰肌醇、磷脂酰乙醇胺和磷脂酰胆碱；响应膳食抗原时，小肠上段中的 CASP3 或 CASP7 会切割 GSDMD 生成转录共激活因子 (依据同源性推测)。该结构域是维持小肠食物耐受的必要成分：它会转位进入细胞核，作为 STAT1 的共激活因子，诱导 CIITA 和 MHC II 类分子的转录，进而诱导小肠上段的 1 型调节性 T (Tr1) 细胞生成 (依据同源性推测)；木瓜蛋白酶过敏原可切割 GSDMD 生成该结构域。切割后，它会转移到质膜，在膜内同源寡聚化，形成内径 10-15 纳米 (nm) 的孔道，使成熟的白细胞介素-33 (IL33) 特异性释放，促进 2 型炎症免疫应答 [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] 。亚细胞定位：细胞质、胞质溶胶；炎症小体；细胞膜；多通道膜蛋白；分泌型；线粒体膜；细胞质、胞质溶胶；细胞核；细胞质、胞质溶胶表达水平：组织特异性：该基因在食管基底上层细胞以及胃峡部/颈部、胃小凹和胃腺中表达，提示其在分化细胞中优先表达。亚基：同源寡聚体；插入膜中时含有 27-28 个亚基的同源寡聚环状孔复合物 (PubMed:33883744, PubMed:34289345, PubMed:38040708)。