Antibodies, Proteins, Kits and Reagents for Life Science

318 288 ₸ Product size

100 µl 318 288 ₸

Order now and get it on Tuesday March 02, 2021

Anti-GSDMD antibody [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, IP, WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat

Product name

Anti-GSDMD antibody [EPR20859]
See all GSDMD primary antibodies
Description

Rabbit monoclonal [EPR20859] to GSDMD
Host species

Rabbit
Specificity

This antibody is recommended for mouse only in IHC.

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
IHC-P	Mouse
IP	Mouse
WB	Mouse Rat

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- 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 tissue. Flow Cyt: RAW 264.7 cells. IP: RAW 264.7 whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20859
Isotype

IgG
Research areas

Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)

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 kDa

Exposure 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.
Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)

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 kDa

Exposure 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, NIBS.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800)

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 our collaborator Dr. Feng Shao NIBS.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.
Flow Cytometry - Anti-GSDMD antibody [EPR20859] (ab219800)

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.
Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (ab219800)

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.
Anti-GSDMD antibody [EPR20859] (ab219800)

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