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Gasdermin D (N terminal) Recombinant Rabbit Monoclonal Antibody [PD00-18]

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Catalog# HA721144

Gasdermin D (N terminal) Recombinant Rabbit Monoclonal Antibody [PD00-18]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

Gasdermin D (N terminal) Recombinant Rabbit Monoclonal Antibody [PD00-18]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Recombinant protein within Gasdermin D full length protein.

种属反应性

Human, Mouse, Rat

验证应用

WB, IHC-P, IF-Cell

分子量

Predicted band size: 53/30 kDa

阳性对照

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.

偶联

unconjugated

克隆号

PD00-18

RRID

AB_3072266

产品特性

形态

Liquid

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

存储缓冲液

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:2,000-1:5,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

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).

亚细胞定位

Cell membrane, Secreted.

UNIPROT

SwissProt: P57764 Human

SwissProt: Q9D8T2 Mouse

Entrez Gene: 513939 Rat

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

图片

  • Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/2,000 dilution.<br /><br />Lane 1: SiHa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: THP-1 cell lysate<br />Lane 4: PC-3M cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: PC-12 cell lysate<br />Lane 7: L6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 53/30 kDa<br />Observed band size: 53/30 kDa<br /><br />Exposure time: 43 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/2,000 dilution.<br /><br />Lane 1: THP-1 cell lysate (20 µg/Lane)<br />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)<br /><br />Predicted band size: 53/30 kDa<br />Observed band size: 53/35 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse intestine tissue with Rabbit anti-Gasdermin D (N terminal) antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse intestine tissue with Rabbit anti-Gasdermin D (N terminal) 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.

  • Immunocytochemistry analysis of L6 cells labeling Gasdermin D (N terminal) with Rabbit anti-Gasdermin D (N terminal) antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/100 dilution.<br /><br />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 (N terminal) antibody (<a href="/products/HA721144" style="font-weight: bold;text-decoration: underline;">HA721144</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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 (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 (N terminal) 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.

请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

引文

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