Recombinant Anti-GSDMD antibody [EPR19828] (ab209845)
![Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-481628-anti-gsdmd-antibody-epr19828-western-blot-mouse-5.JPG "Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19828] to GSDMD
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-GSDMD antibody [EPR19828]
See all GSDMD primary antibodies -
Description
Rabbit monoclonal [EPR19828] to GSDMD -
Host species
Rabbit -
Specificity
In our hands this product works well in western blot using mouse cell lines but not tissue samples. We recommend ab219800 as an alternative for western blotting of mouse tissue samples. -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: RAW 264.7 cell line with ectopic expression of Apoptosis-associated speck-like protein, untreated and primed with 1 µg/mL lipopolysaccharides for 4 h followed by 10 µM nigericin treatment under serum starved conditions for 2 h, whole cell lysates; J774A.1. Mouse liver lysate IP: RAW 264.7-derived cells with ectopic expression of ASC, untreated and 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 lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19828 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab209845 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (2) |
1/1000. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa).
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa). |
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IP
1/30. |
Target
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Sequence similarities
Belongs to the gasdermin family. - Information by UniProt
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Database links
- Entrez Gene: 69146 Mouse
- SwissProt: Q9D8T2 Mouse
- Unigene: 27385 Mouse
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Alternative names
- 1810036L03Rik antibody
- DF 5L antibody
- DF5L antibody
see all
Images
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Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)All lanes : Anti-GSDMD antibody [EPR19828] (ab209845) at 1/1000 dilution (ab219800)
Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 1
Lane 2 : Mouse liver lysate
Lane 3 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 2
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: ab209845 - 120 seconds, ab219800 - 20 seconds
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).
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![Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-320338-anti-gsdmd-antibody-epr19828-western-blot.jpg)
Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)Lanes 1-3 : Anti-GSDMD antibody [EPR19828] (ab209845) at 1/1000 dilution
Lanes 4-6 : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution
Lanes 1 & 4 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lanes 2 & 5 : Mouse liver tissue lysate
Lanes 3 & 6 : Mouse spinal cord tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 3 minutesBlocking/Diluting buffer and concentration 5% NFDM/TBST.
Ab209845 work best with cell lysates.
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![Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-279242-anti-gsdmd-antibody-epr19828-western-blot.jpg)
Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)All lanes : Anti-GSDMD antibody [EPR19828] (ab209845) 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
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
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
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 53 kDa
Observed band size: 32,53 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/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.
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![Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-320340-anti-gsdmd-antibody-epr19828-western-blot.jpg)
Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)Lane 1 : Anti-GSDMD antibody [EPR19828] (ab209845) at 1/1000 dilution
Lane 2 : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution
All lanes : J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 3 minutesDiluting/Blocking buffer and concentration 5% NFDM/TBST
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![Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-279239-anti-gsdmd-antibody-epr19828-immunoprecipitation.jpg)
Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (ab209845)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.
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.
All lanes :
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 : 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 at 10 µg
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 -
![Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-464252-anti-gsdmd-antibody-epr19828-westernblot-mouse.jpg)
Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)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.
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![Anti-GSDMD antibody [EPR19828] (ab209845)](https://www.abcam.com/ps/products/209/ab209845/Images/ab209845-6-benefits-of-recombinant-antibodies.png)
Anti-GSDMD antibody [EPR19828] (ab209845)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (111)
ab209845 has been referenced in 111 publications.
- Liu L et al. Citrobacter freundii Activation of NLRP3 Inflammasome via the Type VI Secretion System. J Infect Dis 223:2174-2185 (2021). PubMed: 33151309
- Chang H et al. Micro-ribonucleic acid-23a-3p prevents the onset of type 2 diabetes mellitus by suppressing the activation of nucleotide-binding oligomerization-like receptor family pyrin domain containing 3 inflammatory bodies-caused pyroptosis through negatively regulating NIMA-related kinase 7. J Diabetes Investig 12:334-345 (2021). PubMed: 32881354
- Muendlein HI et al. ZBP1 promotes LPS-induced cell death and IL-1ß release via RHIM-mediated interactions with RIPK1. Nat Commun 12:86 (2021). PubMed: 33397971
- Estfanous S et al. Gasdermin D restricts Burkholderia cenocepacia infection in vitro and in vivo. Sci Rep 11:855 (2021). PubMed: 33441602
- Wang Y et al. Type 1 interferon aggravates lipopolysaccharide-induced sepsis through upregulating Caspase-11 and Gasdermin D. J Physiol Biochem 77:85-92 (2021). PubMed: 33515436