Anti-GSDMD antibody [EPR19828] - BSA and Azide free
- RabMAb
- Recombinant
- KO Validated
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(7 Publications)
Rabbit Recombinant Monoclonal GSDMD antibody. Carrier free. Suitable for IP, WB and reacts with Mouse samples. Cited in 7 publications.
View Alternative Names
Gsdmdc1, Gsdmd, Gasdermin-D, Gasdermin domain-containing protein 1
- IP
Unknown
Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
Gasdermin-D was immunoprecipitated from 0.35 mg of RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cells with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate with ab209845 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab209845 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate 10ug (Input).
Lane 2 : ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab209845 in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.
Details of RAW 264.7-derived cells with ectopic expression of ASC are described in the literature : PMID 26611636.
The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209845).
All lanes:
Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)
Predicted band size: 53 kDa
false
- IP
Supplier Data
Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
Gasdermin-D was immunoprecipitated from 0.35 mg of RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate with ab209845 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab209845 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate 10ug (Input).
Lane 2 : ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab209845 in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.
As a response to LPS stimulation and nigericin treatment, Gasdermin-D is cleaved and Gasdermin-D N-terminal form is detected at 32kDa. Details of RAW 264.7-derived cells with ectopic expression of ASC are described in the literature : PMID 26611636.
The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.
Note : The antibody has better affinity to full length Gasdermin-D.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209845).
All lanes:
Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a>) at 1/1000 dilution
Lane 1:
RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h at 10 µg
Lane 2:
<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a> IP in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h whole cell lysate at 10 µg
Lane 3:
Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a> in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h whole cell lysate
Predicted band size: 53 kDa
false
- WB
Lab
Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
Blocking and dilution buffer : 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation (ab209845).
All lanes:
Western blot - Anti-GSDMD antibody [EPR19828] (<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a>) at 1/1000 dilution
Lane 1:
J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg
Lane 2:
J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) treated with 100ng/ml lipopolysaccharides (LPS) for 3h, then added 300ng/ml BFA for another 3h whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
Exposure time: 180s
- WB
Lab
Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
This data was developed using ab209845, the same antibody clone in a different buffer formulation. Different batches of ab209845 were tested on :
1 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) -derived cell line with ectopic expression of ASC primed with 1 μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 uM nigericin treatment under serum starved conditions for 2 h, whole cell lysate at 5.2 μg/ml.
2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) -derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate at 5.2 μg/ml.
15 μg of lysate was loaded in each lane. Bands observed at 32,53 kDa.
All lanes:
Western blot - Anti-GSDMD antibody [EPR19828] (<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a>)
Predicted band size: 53 kDa
false
- WB
Supplier Data
Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
Blocking/Dilution buffer : 5% NFDM/TBST.
As a response to LPS stimulation and nigericin treatment, Gasdermin-D is cleaved and Gasdermin-D N-terminal form is detected at 32kDa. Details of RAW264.7-derived cells with ectopic expression of ASC are described in the literature : PMID 26611636
The MW observed is consistent with the literature : PMID 26375003.
The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209845).
All lanes:
Western blot - Anti-GSDMD antibody [EPR19828] (<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a>) at 1/1000 dilution
Lane 1:
RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate at 20 µg
Lane 2:
RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC primed with 1 μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate at 20 µg
Lane 3:
Gasdermin-D knockout RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution
Predicted band size: 53 kDa
Observed band size: 32 kDa,53 kDa
false
Exposure time: 1min
- WB
Lab
Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867)
This data was developed using ab209845, the same antibody clone in a different buffer formulation.
Compared with ab209845, ab219800 has a higher affinity in positive samples.
The MW observed is consistent with the literature : PMID 26375003.
The 43 kDa band is likely to be a GSDMD cleavage product by caspase-3 at GSDMD Asp88 (PMID : 30902848, PMID : 28392147, PMID : 30135078).
We are unsure how to define the 37kDa extra band.
Other bands may be the GSDMD cleavage products, you can find them in PMID : 34890644, PMID : 33208231, PMID : 28679757.
Blocking and dilution buffer : 5% NFDM/TBST.
All lanes:
Western blot - Anti-GSDMD antibody [EPR19828] (<a href='/en-us/products/primary-antibodies/gsdmd-antibody-epr19828-ab209845'>ab209845</a>) at 1/1000 dilution
Lane 1:
RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 1 at 20 µg
Lane 2:
Mouse liver lysate at 20 µg
Lane 3:
NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 20 µg
Lane 4:
RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 2 at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
Exposure time: 120s
Related conjugates and formulations (1)
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Anti-GSDMD antibody [EPR19828]
Reactivity data
Product details
ab225867 is the carrier-free version of ab209845.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties and storage information
Form
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Gasdermin D functions in the execution of immune responses against infections. It acts as an effector molecule that participates directly in pyroptosis by disrupting mitochondrial membranes. GSDMD operates as part of a larger inflammasome complex initiated by inflammatory signals. The inflammasome activates inflammatory caspases that cleave GSDMD enabling its active form to execute pyroptosis. This process releases cytokines like IL-1? enhancing the inflammatory response.
Pathways
GSDMD is important in the pyroptosis pathway initiated by the inflammasome. This process involves Caspase-1 a protease responsible for cleaving pro-inflammatory cytokines and initiating pyroptosis. Another significant pathway includes NLRP3 inflammasome which acts upstream to activate Caspase-1 and subsequently GSDMD establishing the overall inflammatory response in the innate immune system. Through these pathways GSDMD interacts closely with proteins like IL-18 an essential inflammatory mediator.
Product protocols
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Target data
Publications (7)
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Cell death & disease 16:426 PubMed40456737
2025
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International journal of oncology 66: PubMed39950328
2025
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Unspecified reactive species
Functional & integrative genomics 24:55 PubMed38467948
2024
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Advanced science (Weinheim, Baden-Wurttemberg, Germany) 11:e2303177 PubMed38308188
2024
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Gut microbes 15:2180315 PubMed36803521
2023
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Unspecified reactive species
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 41:376-383 PubMed33849828
2021
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Unspecified reactive species
Toxicology and applied pharmacology 377:114626 PubMed31201821
2019
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Product promise
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