Anti-GSDMD 抗体 [EPR20859] (ab219800)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20859] to GSDMD
- Suitable for: Flow Cyt (Intra), IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat
Related conjugates and formulations
製品の概要
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製品名
Anti-GSDMD antibody [EPR20859]
GSDMD 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR20859] to GSDMD -
由来種
Rabbit -
特異性
This antibody can detect full length, as well as a N-terminal fragment after stimulation in WB.
Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487), we recommend loading higher amount of lysate or using lower antibody dilution.
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アプリケーション
適用あり: Flow Cyt (Intra), IP, WB, IHC-Pmore details
適用なし: ICC/IF -
種交差性
交差種: Mouse, Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Wild-type mouse liver and lung whole cell lysates; C6, PC-12, RAW 264.7 and NIH/3T3 whole cell lysates; Rat brain and liver lysates, IHC-P: Wild type mouse small intestine and mouse lung tissues. Flow Cyt (intra): RAW 264.7 cells. IP: RAW 264.7 whole cell lysate.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR20859 -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab219800の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/500.
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IP |
1/30.
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WB |
1/1000 - 1/2000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
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IHC-P | (2) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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特記事項 |
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Flow Cyt (Intra)
1/500. |
IP
1/30. |
WB
1/1000 - 1/2000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ターゲット情報
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配列類似性
Belongs to the gasdermin family. - Information by UniProt
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参照データベース
- Entrez Gene: 69146 Mouse
- Entrez Gene: 315084 Rat
- SwissProt: Q9D8T2 Mouse
- Unigene: 27385 Mouse
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別名
- 1810036L03Rik antibody
- DF 5L antibody
- DF5L antibody
see all
画像
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Immunohistochemical analysis of paraffin-embedded (A) Small intestine tissue from wild-type C57BL/6JGpt mice (B) Small intestine tissue from GSDMD knockout mice staining with ab219800 at 1/5000 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Small intestine tissue from wild-type C57BL/6JGpt mice and no staining on (B) Small intestine tissue from GSDMD knockout mice. The section was incubated with ab219800 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and GSDMD-KO homozygous mice (Strain ID: T010437).
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue labelling GSDMD with ab219800 at 1/1000 (0.542 μg/ml) followed by ab214880 at a ready to use dilution. Low expression tissue: weak staining on mouse lung. The section was incubated with ab219800 at 4°C overnight. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is ab214880 at a ready to use dilution.
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lane 5 : Rat liver lysate
Lane 6 : Rat brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaExposure time : Lanes 1 and 2:3 minutes; Lanes 3 and 4:48 seconds; Lanes 5 and 6:3 minutes.
Blocking/Dilution buffer: 5% NFDM/TBST.
Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487), we recommend loading higher amount of lysate or using lower antibody dilution.
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All lanes : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution
Lane 1 : Wild-type mouse liver whole cell lysate
Lane 2 : GSDMD knockout mouse liver whole cell lysate
Lane 3 : Wild-type mouse lung whole cell lysate
Lane 4 : GSDMD knockout mouse lung whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaExposure time: Lanes 1 and 2:15 seconds; Lanes 3 and 4:37 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The antibody has strong reactivity with GSDMD with a few weak bands of undermined proteins.
The lysates were kindly provided by our collaborator Dr Feng Shao's lab, NIBS.
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Immunohistochemical analysis of paraffin-embedded Wild type mouse small intestine (A) and GSDMD KO mouse small intestine (B) tissue labeling GSDMD with ab219800 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on epithelium of wild type mouse small intestine (A), no staining on GSDMD KO mouse small intestine (B) (PMID: 17350798) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
The tissue samples were kindly provided by Dr. Feng Shao NIBS.
Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.
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All lanes : Anti-GSDMD antibody [EPR20859] (ab219800)
Lane 1 : iBMM (mouse immortalized bone marrow derived macrophages) treated with 500ng/ml Bsak plus 500ng/ml anthrax protective antigen (PA) for 2h. Cell lysate
Lane 2 : iBMM (mouse immortalized bone marrow derived macrophages) treated with 500ng/ml Bsak plus 500ng/ml anthrax protective antigen (PA) for 2h. Concentrated cell supernatant
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 31,53 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking and diluting buffer: 5% NFDM /TBST
This antibody could detect full length, as well as a N-terminal fragment after stimulation.The lysates were kindly provided by our Dr Feng Shao's lab, NIBS. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling GSDMD with ab219800 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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GSDMD was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate with ab219800 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219800 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: RAW 264.7 whole cell lysate 10 µg (Input).
Lane 2: ab219800 IP in RAW 264.7 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219800 in RAW 264.7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (96)
ab219800 は 96 報の論文で使用されています。
- Tojo K et al. Early alveolar epithelial cell necrosis is a potential driver of COVID-19-induced acute respiratory distress syndrome. iScience 26:105748 (2023). PubMed: 36507222
- Liu Q et al. Serum IL-1, Pyroptosis and Intracranial Aneurysm Wall Enhancement: Analysis Integrating Radiology, Serum Cytokines and Histology. Front Cardiovasc Med 9:818789 (2022). PubMed: 35155635
- Cao Z et al. Crosstalk of pyroptosis, ferroptosis, and mitochondrial aldehyde dehydrogenase 2-related mechanisms in sepsis-induced lung injury in a mouse model. Bioengineered 13:4810-4820 (2022). PubMed: 35188436
- Zhang T et al. CircNEIL3 mediates pyroptosis to influence lung adenocarcinoma radiotherapy by upregulating PIF1 through miR-1184 inhibition. Cell Death Dis 13:167 (2022). PubMed: 35190532
- Yan J et al. Metformin reduces chondrocyte pyroptosis in an osteoarthritis mouse model by inhibiting NLRP3 inflammasome activation. Exp Ther Med 23:222 (2022). PubMed: 35222699