Protein extraction was performed using Ripa Buffer and Protease/Phosphatase Inhibitor Cocktail (Cell Signaling) and protein quantification using the Bradford method. Protein samples (30 μg) were mixed 1:1 with Laemmli buffer supplemented with 20% 1,4-Dithiothreitol (DTT) (Sigma) and loaded on 4–12% Tris-Glycine gels (Lonza). Running was performed at 125 V for 80′ and wet transfer at 25 V for 120′. Membranes were blocked with 5% milk in TBS-t for 1 h, at room temperature (RT). The primary Abs used were: anti-caspase-1 (2225, Cell Signaling), anti-ASC (AG-25B-0006, Adipogen) and anti-β-tubulin (2128, Cell Signaling). The secondary Abs were: peroxidase goat anti-rabbit IgG (H + L) (PI-1000) and peroxidase horse anti-mouse IgG (H + L) (PI-2000, Vector Laboratories). For detection we used ECL Select Reagent (Amersham, GE Healthcare), ChemiDoc™ MP system and Image Lab Software (Bio-Rad).
Anti asc ag 25b 0006
Manufactured by Adipogen
Sourced in United States
Anti-ASC (AG-25B-0006) is a primary antibody that recognizes the Apoptosis-associated Speck-like protein containing a CARD (ASC). ASC is an important component of the inflammasome, a multi-protein complex involved in the activation of inflammatory caspases. This antibody is useful for the detection of ASC in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Anti caspase 1, by Cell Signaling Technology (1 mentions)
Ecl select reagent, by Cytiva (1 mentions)
Anti β tubulin, by Cell Signaling Technology (1 mentions)
Protease phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
1 4 dithiothreitol dtt, by Merck Group (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Chemidoc mp system, by Bio-Rad (1 mentions)
Nigericin, by Bio-Techne (1 mentions)
Anti β actin antibody sc 47778, by Santa Cruz Biotechnology (1 mentions)
Salmonella typhimurium flagellin, by InvivoGen (1 mentions)
Anti il 1β antibody af 401 na, by R&D Systems (1 mentions)
Poly da dt, by InvivoGen (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Sinapic acid, by Merck Group (1 mentions)
Nigericin, by InvivoGen (1 mentions)
Selisistat ex 527, by MedChemExpress (1 mentions)
Alexa fluor 488 conjugated anti rabbit and, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti asc ag 25b 0006
1
Western Blot Protein Analysis Protocol
2
Inflammatory Responses Mediated by NLRP3
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Sinapic acid (SA, 3,5-dimethoxy-4-hydroxycinnamic acid), LPS, and ATP were purchased from Sigma-Aldrich (St. Louis, MO, USA). Nigericin was purchased from Tocris (Bristol, UK). Poly (dA:dT) and Salmonellatyphimurium flagellin were purchased from InvivoGen (San Diego, CA, USA). Anti-IL-1β antibody (AF-401-NA) was purchased from R&D Systems (Minneapolis, MN, USA). The anti-caspase-1 (AG-20B-0042), anti-NLRP3 (AG-20B-0014), and anti-ASC (AG-25B-0006) antibodies were purchased from AdipoGen Life Science (San Diego, CA, USA). Anti-IL-6 (12912), anti-phospho-STAT3 (9145), anti-phospho-IκBα (9246), anti-phospho-JNK (4668), and anti-phospho-ERK (9106) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-β-actin antibody (sc-47778) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). All culture reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
3
Melatonin Neuroprotection in MPTP Model
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Melatonin (M5250), MPTP-HCL (M0896), 1-methyl-4-phenylpyridinium (MPP+, D048), adenosine 5-triphosphate disodium hydrate (ATP, A6419) were obtained from Sigma‒Aldrich (St Louis, MO), and nigericin was purchased from InvivoGen (San Diego, CA). Selisistat (EX-527) was purchased from MedChemExpress (Monmouth Junction, NJ). Anti-NLRP3 (AG-20B-0014), anti-caspase1 (AG-20B-0042), and anti-ASC (AG-25B-0006) antibodies were purchased from AdipoGen Life Sciences (San Diego, CA). Anti-IBA-1 (ab5076) antibody and anti-NF-κB p65 (ab16502) antibody were purchased from Abcam (Cambridge, MA). Anti-tyrosine hydroxylase (TH) (AB152) was purchased from Merck Millipore (Billerica, MA). Anti-SIRT1 (No.8469) antibody and anti-phospho-NF-κB p65 (No.3033) were purchased from Cell Signaling Technology (Danvers, MA). Anti-IL18 (No. A1115) antibody was purchased from ABclonal (Woburn, MA). Alexa fluor-488 conjugated anti-rabbit and Alexa fluor-546 conjugated anti-goat antibodies were purchased from Invitrogen (Carlsbad, CA). The mouse IL-1β ELISA kit (EK0394) was purchased from Boster Bio (Pleasanton, CA). The 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe was purchased from NJJCBIO (Nanjing, China).
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