- PA5-116815 - Invitrogen Antibodies GSDMD antibody

Xu J, Wang Q, Song YF, Xu XH, Zhu H, Chen PD, Ren YP
World journal of diabetes 2022 Apr 15;13(4):358-375

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of GSDMD using GSDMD Polyclonal Antibody (Product # PA5-116815) at a 1:1,000 dilution. THP-1 cells were treated by PMA/TPA (80 nM) at 37°C for overnight and LPS (1 µg/mL) at 37°C for 6 hours. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10,000 dilution. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry/Immunofluorescence analysis of GSDMD in BALB-3T3 cells using GSDMD Polyclonal Antibody (Product # PA5-116815) at a dilution of 1:100. Blue: DAPI for nuclear staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry/Immunofluorescence analysis of GSDMD in C6 cells using GSDMD Polyclonal Antibody (Product # PA5-116815) at a dilution of 1:100. Blue: DAPI for nuclear staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry/Immunofluorescence analysis of GSDMD in U-2 OS cells using GSDMD Polyclonal Antibody (Product # PA5-116815) at a dilution of 1:100. Blue: DAPI for nuclear staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of GSDMD in rat spleen tissue using GSDMD Polyclonal Antibody (Product # PA5-116815) at a dilution of 1:100.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation analysis of GSDMD in 300 µg extracts of Jurkat cells using GSDMD Polyclonal Antibody (Product # PA5-116815). Western blot was performed from the immunoprecipitate using the same antibody at a dilution of 1:500

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation analysis of GSDMD in 300 µg extracts of Jurkat cells using GSDMD Polyclonal Antibody (Product # PA5-116815). Western blot was performed from the immunoprecipitate using the same antibody at a dilution of 1:500

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Silencing lncRNA X inactive specific transcript can inhibit renal tubular epithelial cell pyroptosis in diabetic nephropathy rats. After establishing diabetic nephropathy model rats, lentivirus-LV sh- X inactive specific transcript was injected into the tail vein. A: Enzyme-linked immunosorbent assay detected IL-1beta and IL-18 expressions; B: Western blot tested the levels of NLR family pyrin domain containing 3, ASC, Cleaved Caspase-1, Caspase-1, GSDMD, and GSDMD-N; C: Immunofluorescence tested the expression of Caspase-1; scale bar: 25 mum; N = 8/group. The data were described as mean +- SD and analyzed by one-way ANOVA and Tukey's multiple comparisons test; a P < 0.05. DN: Diabetic nephropathy; LV: Lentivirus; XIST: X inactive specific transcript; sh: Short hairpin RNA; NLRP3: NLR family pyrin domain containing 3; ASC: Apoptosis speck-like protein; GSDMD: Gasdermin D; DAPI: 4',6-diamidino-2-phenylindole.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Silencing X inactive specific transcript in diabetic nephropathy can inhibit renal tubular epithelial cell pyroptosis in high glucose-induced HK2 cells. si-NC or si- X inactive specific transcript (XIST) was delivered into HK2 cells treated with high glucose. A: The XIST expression detected by qRT-PCR; B: Enzyme-linked immunosorbent assay detected the expression of IL-1beta and IL-18; C: Western blot tested the levels of NLR family pyrin domain containing 3, ASC, Cleaved Caspase-1, Caspase-1, GSDMD, and GSDMD-N; D: Cell viability; E: Lactate dehydrogenase activity. The cell experiment was performed in triplicate. The data were described as mean +- SD and analyzed by one-way ANOVA (A/D-E) or two-way ANOVA (B/C) and Tukey's multiple comparisons test; a P < 0.05. NG: Normal glucose; HG: High glucose; si: Small interfering RNA; XIST: X inactive specific transcript; LDH: Lactate dehydrogenase; NLRP3: NLR family pyrin domain containing 3; SC: Apoptosis speck-like protein; GSDMD: Gasdermin D.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Inhibition of miR-15b-5p in X inactive specific transcript -silenced HK2 cells activates renal tubular epithelial cell pyroptosis. In HG-treated HK2 cells, both X inactive specific transcript (XIST) and miR-15b-5p were silenced. A: qRT-PCR verified the transfection efficiency of miR-15b-5p inhibitor; B: Enzyme-linked immunosorbent assay detected the expression of IL-1beta and IL-18; C: Western blot tested the levels of NLR family pyrin domain containing 3, ASC, Cleaved Caspase-1, Caspase-1, GSDMD, and GSDMD-N; D: Cell viability; E: Lactate dehydrogenase activity. The cell experiment was performed in triplicate. The data were described as mean +- SD and analyzed by one-way ANOVA (A/D-E) or two-way ANOVA (B/C) and Tukey's multiple comparisons test; a P < 0.05. LDH: Lactate dehydrogenase; XIST: X inactive specific transcript; si: Small interfering RNA; NLRP3: NLR family pyrin domain containing 3; ASC: Apoptosis speck-like protein; GSDMD: Gasdermin D.

PA5-116815