Gasdermin D Recombinant Rabbit Monoclonal Antibody [PD00-18]
cat.: HA750486
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, IF-Tissue
Clonality: Monoclonal
Clone number: PD00-18
Form: Liquid
Storage condition: Store at 2-8℃. Avoid freeze.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 53/30 kDa
Isotype: IgG
Immunogen: Recombinant protein within Gasdermin D full length protein.
Positive control: SiHa cell lysate, Jurkat cell lysate, THP-1 cell lysate, PC-3M cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, L6 cell lysate, THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate, mouse intestine tissue, L6.
Subcellular location: Cell membrane, Secreted.
Uniprot #: SwissProt: P57764 Human | Q9D8T2 Mouse
Entrez Gene: 513939 Rat
Alternative names: 1810036L03Rik DF 5L DF5L DFNA 5L DFNA5L FKSG 10 FKSG10 FLJ12150 Gasdermin Gasdermin D Gasdermin domain containing 1 Gasdermin domain containing protein 1 Gasdermin domain-containing protein 1 Gasdermin-D GasderminD GSDMD GSDMD_HUMAN GSDMDC 1 GSDMDC1
Images
HA750486_1.jpg Fig1: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA750486) at 1/2,000 dilution.

Lane 1: SiHa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: THP-1 cell lysate
Lane 4: PC-3M cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: PC-12 cell lysate
Lane 7: L6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 53/30 kDa
Observed band size: 53/30 kDa

Exposure time: 43 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750486) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA750486_2.jpg Fig2: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA750486) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate (20 µg/Lane)

Predicted band size: 53/30 kDa
Observed band size: 53/35 kDa

Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750486) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA750486_3.jpg Fig3: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse intestine tissue with Rabbit anti-Gasdermin D antibody (HA750486) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750486) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA750486_4.jpg Fig4: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of L6 cells labeling Gasdermin D with Rabbit anti-Gasdermin D antibody (HA750486) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Gasdermin D antibody (HA750486) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA750486_5.jpg Fig5: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Application: Immunofluorescence (IF-tissue)

Species: Mouse
Tissue: Intestine
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA750486, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA750486_6.jpg Fig6: This data was developed using HA750486, the same antibody clone in a different buffer formulation.
Western blot analysis of Gasdermin D on different lysates with Rabbit anti-Gasdermin D antibody (HA750486) at 1/2,000 dilution.

Lane 1: HCT-8 WT
Lane 2: HCT-8 GSDMD KO
Lane 3: HCT-8 GSDMD KO+empty vector
Lane 4: HCT-8 GSDMD KO+OE-GSDMD

Lysates/proteins at 20 µg/Lane.
Exposure time: 59 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA750486, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 53 kDa
Observed band size: 53 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.