GSDMD Recombinant monoclonal antibody

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  WB analysis of NIH/3T3 using 83562-3-RR

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 83562-3-RR (Gsdmd antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis using 83562-3-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83562-3-RR (Gsdmd antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IF Staining of NIH/3T3 using 83562-3-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed NIH/3T3 cells using Gsdmd antibody (83562-3-RR, Clone: 240548B10 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).

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  FC experiment of RAW 264.7 using 83562-3-RR

  1x10^6 RAW 264.7 cells were intracellularly stained with 0.25 ug Gsdmd Recombinant antibody (83562-3-RR, Clone:240548B10) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

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  Affinity and Kinetic Characterization of 83562-3-RR

  Biolayer interferometry (BLl) kinetic assays of 83562-3-RR against Mouse Gsdmd were performed. The affinity constant is 1.19 nM.

GSDMD Uni-rAb^® Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Cat No. 83562-3-RR

Clone No.240548B10

Host / Isotype

Rabbit / IgG

Reactivity

mouse

Applications

WB, IF/ICC, FC (Intra), ELISA

Synonyms

240548B10, AW558049, DF5L, Dfna5l, gasdermin D

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Tested Applications

• Positive WB detected in: NIH/3T3 cells, EL-4 cells, RAW 264.7 cells, J774A.1 cells
• Positive IF/ICC detected in: NIH/3T3 cells
• Positive FC (Intra) detected in: RAW 264.7 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

83562-3-RR targets GSDMD in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with mouse samples.

• Tested Reactivity: mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  CatNo: Ag34905

  Product name: Recombinant mouse Gsdmd protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 199-487 aa of BC029813

  Sequence: GHQSRKKMVTIPAGSILAFRVAQLLIGSKWDILLVSDEKQRTFEPSSGDRKAVGQRHHGLNVLAALCSIGKQLSLLSDGIDEEELIEAADFQGLYAEVKACSSELESLEMELRQQILVNIGKILQDQPSMEALEASLGQGLCSGGQVEPLDGPAGCILECLVLDSGELVPELAAPIFYLLGALAVLSETQQQLLAKALETTVLSKQLELVKHVLEQSTPWQEQSSVSLPTVLLGDCWDEKNPTWVLLEECGLRLQVESPQVHWEPTSLIPTSALYASLFLLSSLGQKPC*

• Full Name: gasdermin D
• Calculated Molecular Weight: 53 kDa
• Observed Molecular Weight: 53 kDa
• GenBank Accession Number: BC029813
• Gene Symbol: Gsdmd
• Gene ID (NCBI): 69146
• RRID: AB_3671177
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: Q9D8T2
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

GSDMD is a 53 kDa cytosolic protein and a member of the gasdermin protein family, which features 6 members in humans (GSDMA, -B, -C, -D, -E and -F. GSDMD was identified as the executioner of inflammasome-associated pyroptosis. Caspase-1, -11, or -4 cleave GSDMD after an aspartate residue within a central linker domain (D275 in humans and D276 in mice) to generate a 31-kDa N-terminal GSDMD fragment (GSDMD-NT) and a 22-kDa C-terminal fragment (GSDMD-CT). In addition to the canonical 31/22 kDa cleavage products, a 43 kDa fragment has been observed under specific experimental or pathological conditions. The generation of the 43 kDa fragment was observed upon caspase-3 and -7 activation during apoptosis. This antibody can only detect full-length GSDMD.