Anti goat igg hrp linked antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-goat IgG HRP-linked Antibody is a detection reagent used in immunoassays and Western blot analyses. It is an antibody directed against goat immunoglobulin G (IgG) that is conjugated to the enzyme horseradish peroxidase (HRP). The HRP label allows for the detection and visualization of target proteins bound by the primary antibody.

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HRP-anti-goat IgG (2 citations) Donkey-anti-goat IgG HRP-linked antibody (2 citations)

3 protocols using anti goat igg hrp linked antibody

Western Blot Analysis of Signaling Proteins

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The Western blot experiments were done as described previously [52 (link)]. Mouse monoclonal antibodies, at concentrations recommended by the manufacturer, were used to detect A20 (Santa Cruz Biotechnology, Dallas, TX, USA), NF-κB p65 (Santa Cruz Biotechnology) and phosphorylated (p)-IκBα (Cell Signaling Technology, Danvers, MA, USA) and a goat polyclonal antibody was used to detect IκBα (Santa Cruz Biotechnology). As secondary antibody, an anti-mouse IgG horseradish peroxidase (HRP)-linked antibody (Santa Cruz Biotechnology) or anti-goat IgG HRP-linked antibody (Santa Cruz Biotechnology) was used.

Rigauts C., Aizawa J., Taylor S.L., Rogers G.B., Govaerts M., Cos P., Ostyn L., Sims S., Vandeplassche E., Sze M., Dondelinger Y., Vereecke L., Van Acker H., Simpson J.L., Burr L., Willems A., Tunney M.M., Cigana C., Bragonzi A., Coenye T, & Crabbé A. (2022). Rothia mucilaginosa is an anti-inflammatory bacterium in the respiratory tract of patients with chronic lung disease. The European Respiratory Journal, 59(5), 2101293.

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Western Blot Analysis of ER Stress Markers

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Whole protein extracts were recovered from cells after treatment with DMSO or SIX2G by using NP40 lysis buffer complemented with Halt Protease and Phosphatase Inhibitor cocktail (Invitrogen by Thermo Fisher Scientific, Waltham, MA, USA). Western blot (WB) analysis was performed as previously reported [26 (link)]. The primary antibodies used were: PERK (#3192), eIF2α (#5324), p-eIF2α (Ser 51) (#3398), Caspase-3 (#9662), Cleaved Caspase-3 (Asp 175) (#9661), Caspase-8 (#4790) and Cleaved Caspase-8 (#9748) by Cell Signaling Technology (Danvers, MA, USA); p-PERK (Thr 981) (#orb336657), b-actin (#Sc-1616), GAPDH (SC-25778), GADD34 (SC-46661) and PP1 (SC-7482) by Santa-Cruz (Dallas, TX, USA). Anti-rabbit IgG HRP-linked antibody (#7074, Cell Signaling Technology, Danvers, MA, USA), anti-mouse IgG HRP-linked antibody (#7076, Cell Signaling Technology, Danvers, MA, USA) and anti-goat IgG HRP-linked antibody (SC-2354, Santa Cruz, Dallas, TX, USA) were used as secondary antibodies depending on the host species of animal in which the primary antibodies were raised.

Grillone K., Riillo C., Rocca R., Ascrizzi S., Spanò V., Scionti F., Polerà N., Maruca A., Barreca M., Juli G., Arbitrio M., Di Martino M.T., Caracciolo D., Tagliaferri P., Alcaro S., Montalbano A., Barraja P, & Tassone P. (2022). The New Microtubule-Targeting Agent SIX2G Induces Immunogenic Cell Death in Multiple Myeloma. International Journal of Molecular Sciences, 23(18), 10222.

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Western Blot Analysis of Inflammatory Markers

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Lung tissues were snap‐frozen and protein was isolated. For caspase‐1 immunoblotting, semi‐dry membrane transfer was performed. Anti‐caspase‐1 (14F468) 1:200 (sc‐56036; Santa Cruz Biotechnology Inc.) and anti‐GAPDH 1:30,000 were used as primary antibodies, and rabbit anti‐mouse HRP 1:2500 in 5% fat‐free milk in TBST (P0260; Dako) as the secondary antibody. For IL‐1β and IL‐18 immunoblotting, wet membrane transfer was performed. Anti‐IL‐1β (1:1000) (IL‐1β IL‐F2 NBP1‐42767; Novus Biologicals) and anti‐IL‐18 (1 µg/ml) (IL‐18/IL‐1F4, AF521; R&D Systems) were used as primary antibodies and anti‐rabbit IgG HRP (#1706515; Bio‐Rad Laboratories Inc.) or anti‐goat IgG HRP‐linked antibody (1:2000) (sc‐2020; Santa Cruz Biotechnology Inc.) as secondary antibodies, respectively. All antibodies were diluted in 2.5% fat‐free milk in TBS‐T. FIJI‐ImageJ (NIH) was used for the quantification of the immunoblots. The cleavage ratio for caspase‐1, IL‐1β, and IL‐18 was calculated as cleaved caspase‐1 (p20) divided by pro‐caspase‐1 (p45), or cleaved IL‐1β (p15) divided by pro‐IL‐1β (p34), or cleaved IL‐18 (p18) divided by pro‐IL‐18 (p24).

Mavrogiannis E., Hagdorn Q.A., Bazioti V., Douwes J.M., Van Der Feen D.E., Oberdorf‐Maass S.U., Westerterp M, & Berger R.M. (2022). Pirfenidone ameliorates pulmonary arterial pressure and neointimal remodeling in experimental pulmonary arterial hypertension by suppressing NLRP3 inflammasome activation. Pulmonary Circulation, 12(3), e12101.

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