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Anti caspase 1 p10 sc 514

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Anti-caspase-1 p10 (sc-514) is a laboratory reagent produced by Santa Cruz Biotechnology. It is an antibody that recognizes the p10 subunit of the caspase-1 protein.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Anti mouse igg hrp, by Bio-Rad (2 mentions) Anti flag m2 f1804, by Merck Group (2 mentions) Anti rabbit igg hrp, by Bio-Rad (2 mentions) Pmycb, by Santa Cruz Biotechnology (2 mentions) Anti c myc antibodies, by Enzo Life Sciences (2 mentions) Anti rabbit pan actin polyclonal antibody, by Cell Signaling Technology (2 mentions) Supersignal west pico, by Thermo Fisher Scientific (1 mentions) Odyssey fc, by LI COR (1 mentions) Goat anti mouse hrp, by Bio-Rad (1 mentions) Anti flag m2, by Merck Group (1 mentions) Ab85550, by Abcam (1 mentions) Anti vdac, by Cell Signaling Technology (1 mentions) Adi spa 860 f, by Enzo Life Sciences (1 mentions) Cryo 2 ag 20b 0014, by Adipogen (1 mentions) Ab25631, by Abcam (1 mentions) Goat anti rat hrp, by Santa Cruz Biotechnology (1 mentions) Anti caspase 3 8g10 9665, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Caspase-1 (137 citations) Anti-caspase-1 (60 citations) Sc-514 (27 citations) Caspase-1 p10 (11 citations) Anti-caspase-1 p10 (9 citations) Caspase-1 (sc-514) (4 citations) Anti-mouse caspase-1 p10 antibody (sc-514; (3 citations) Anti-caspase-1 (sc-514, (2 citations)

3 protocols using anti caspase 1 p10 sc 514

1

Analyzing NLRC4 Inflammasome Activation in HEK293 Cells

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NLRC4 activity was analyzed in HEK293-cells. Wild-type human NLRC4 was cloned into pMycB (Santa Cruz Biotech) and verified by Sanger sequencing. The c.1022 T->C mutation was introduced by site-directed mutagenesis (Stragene QuikChange) and verified by Sanger sequencing. Mutated and wildtype myc-NLRC4 were transiently transfected into HEK293 cells along with N-terminal FLAG-human pro-caspase-1 and GFP-tagged human ASC using Lipofectamine2000 (Invitrogen). NLRC4 activity was visually measured 30h after transfection by spontaneous formation of GFP-ASC foci (as visualized in live cells by epifluorescent microscopy). Manual enumeration of ASC foci+ was performed over 20 representative fields at 20x magnification. Pro-caspase-1 p45 autoproteolysis was measured directly by western blotting for p35 (anti-FLAG M2 (F1804), 1:1000; Sigma) and p10 fragments (anti-caspase-1 p10 (sc-514), 1:200; Santa Cruz Biotech), using anti-mouse IgG-HRP or anti-rabbit IgG-HRP (Biorad) and enhanced chemiluminescence. The presence of NLRC4 and actin in lysates was confirmed by blotting with anti- c-Myc antibodies (9E10, 1:1000; Enzo Life Sciences) or anti-rabbit pan-actin polyclonal antibody (#4970, 1:5000; Cell Signaling Technology), respectively.
Romberg N., Al Moussawi K., Nelson-Williams C., Stiegler A.L., Loring E., Choi M., Overton J., Meffre E., Khokha M.K., Huttner A.J., West B., Podoltsev N.A., Boggon T.J., Kazmierczak B.I, & Lifton R.P. (2014). Mutation of NLRC4 causes a syndrome of enterocolitis and autoinflammation. Nature genetics, 46(10), 1135-1139.
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2

Protein Complex Detection by Immunoblotting

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As indicated, membranes were blotted with anti-caspase-1 p10 (sc-514, Santa Cruz Biotechnology), anti-caspase-1 p20 (Casper-1, AG-20B-0042, Adipogen), anti-NLRP3 (Cryo-2, AG-20B-0014, Adipogen), anti-ASC (AL177, AG-25B-0006, Adipogen), anti-GAPDH (6C5, CB1001, Calbiochem), anti-DDK (4C5, TA50011, Origene), anti-FLAG (M2, Sigma), anti-His (Tetra-His, Qiagen), anti-caspase-3 (8G10, 9665, Cell Signaling), anti-caspase-8 (1G12, ALX-804-447, Enzo Life Sciences), anti-caspase-11 (17D9, 14340, Cell Signaling), anti-Prohibitin (PA5-27329, Pierce), anti-VDAC (4866, Cell Signaling), anti-TSPO (9530, Cell Signaling), anti-PMP70 (ab85550, Abcam), anti-LAMP2 (ab25631, Abcam), anti-calnexin (ADI-SPA-860-F, Enzo Life Sciences), and anti-GM130 (clone 35/GM130, BD Biosciences) and probed with secondary antibodies donkey anti-rabbit-HRP (NA934V, GE Healthcare), goat anti-mouse-HRP (170-6515, BioRad), goat anti-rat-HRP (sc-2006, Santa Cruz), or rabbit anti-goat-HRP (611520, Invitrogen). Immunoblots were incubated with chemiluminescent substrates SuperSignal West Pico or Femto (Thermo Scientific) and images were captured using an Odyssey Fc (LI-COR Biosciences) imaging device.
Elliott E.I., Miller A.N., Banoth B., Iyer S.S., Stotland A., Weiss J.P., Gottlieb R.A., Sutterwala F.S, & Cassel S.L. (2018). Mitochondrial assembly of the NLRP3 inflammasome complex is initiated at priming. Journal of immunology (Baltimore, Md. : 1950), 200(9), 3047-3052.
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3

Analyzing NLRC4 Inflammasome Activation in HEK293 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
NLRC4 activity was analyzed in HEK293-cells. Wild-type human NLRC4 was cloned into pMycB (Santa Cruz Biotech) and verified by Sanger sequencing. The c.1022 T->C mutation was introduced by site-directed mutagenesis (Stragene QuikChange) and verified by Sanger sequencing. Mutated and wildtype myc-NLRC4 were transiently transfected into HEK293 cells along with N-terminal FLAG-human pro-caspase-1 and GFP-tagged human ASC using Lipofectamine2000 (Invitrogen). NLRC4 activity was visually measured 30h after transfection by spontaneous formation of GFP-ASC foci (as visualized in live cells by epifluorescent microscopy). Manual enumeration of ASC foci+ was performed over 20 representative fields at 20x magnification. Pro-caspase-1 p45 autoproteolysis was measured directly by western blotting for p35 (anti-FLAG M2 (F1804), 1:1000; Sigma) and p10 fragments (anti-caspase-1 p10 (sc-514), 1:200; Santa Cruz Biotech), using anti-mouse IgG-HRP or anti-rabbit IgG-HRP (Biorad) and enhanced chemiluminescence. The presence of NLRC4 and actin in lysates was confirmed by blotting with anti- c-Myc antibodies (9E10, 1:1000; Enzo Life Sciences) or anti-rabbit pan-actin polyclonal antibody (#4970, 1:5000; Cell Signaling Technology), respectively.
Romberg N., Al Moussawi K., Nelson-Williams C., Stiegler A.L., Loring E., Choi M., Overton J., Meffre E., Khokha M.K., Huttner A.J., West B., Podoltsev N.A., Boggon T.J., Kazmierczak B.I, & Lifton R.P. (2014). Mutation of NLRC4 causes a syndrome of enterocolitis and autoinflammation. Nature genetics, 46(10), 1135-1139.
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