Recombinant Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)
![Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)](https://www.abcam.com/ps/products/225/ab225867/Images/ab225867-321126-anti-gsdmd-antibody-epr19828-immunoprecipitation.jpg "Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)")
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 - BSA and Azide free
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-GSDMD antibody [EPR19828] - BSA and Azide free
See all GSDMD primary antibodies -
Description
Rabbit monoclonal [EPR19828] to GSDMD - BSA and Azide free -
Host species
Rabbit -
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-derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), 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
ab225867 is the carrier-free version of ab209845.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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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 ab225867 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 |
Use at an assay dependent concentration. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa).
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| IP |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa). |
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IP
Use at an assay dependent concentration. |
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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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).
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![Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)](https://www.abcam.com/ps/products/225/ab225867/Images/ab225867-321125-anti-gsdmd-antibody-epr19828-immunoprecipitation.jpg)
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).
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![Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)](https://www.abcam.com/ps/products/225/ab225867/Images/ab225867-464253-anti-gsdmd-antibody-epr19828-westernblot-mouse.jpg)
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.
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![Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)](https://www.abcam.com/ps/products/225/ab225867/Images/ab225867-286690-anti-gsdmd-antibody-epr19828-western-blot.jpg)
Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209845).
-
![Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)](https://www.abcam.com/ps/products/225/ab225867/Images/ab225867-4-benefits-of-recombinant-antibodies.png)
Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab225867 has been referenced in 1 publication.
- Wu YL et al. Microcystin-LR promotes necroptosis in primary mouse hepatocytes by overproducing reactive oxygen species. Toxicol Appl Pharmacol 377:114626 (2019). PubMed: 31201821
