The cells were harvested and treated with PRO-PREP (iNtRON BIOTECHNOLOGY, 17081) for 30 min at 4 °C. The lysed cells were centrifuged at 13,000 rpm for 20 min at 4 °C, and protein concentration was calculated using Bicinchoninic Acid Assay Kit (Thermo Fisher Scientific, Waltham, 23227, MA, USA). The proteins were separated using 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). The membrane was blocked with 5% skim milk in TBST for 40 min, and then treated with primary antibody and incubated overnight at 4 °C. For the primary antibodies, BAX (1:1000, Cell Signaling Technology, #2772S), NIX (1:1000, Cell Signaling Technology, #12396) and β-actin (1:5000, sc-47778) were used. Primary antibodies were detected using goat anti-mouse IgG-HRP (1:5000, Enzo Life Sciences, ADI-SAB-100-J, USA) or goat anti-rabbit IgG-HRP (1:5000, Enzo Life Sciences, AD I-SAB-300-J, USA). After the membrane was treated with Immobilon Crescendo Western HRP substrate (Millipore, WBLUR0500, USA), proteins were detected using an X-ray film or confirmed using an Amersham Imager 680 (GE Healthcare).
Goat anti rabbit igg hrp
Manufactured by Enzo Life Sciences
Sourced in United States
Goat anti-rabbit IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and quantify rabbit immunoglobulin G (IgG) in various immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Amersham imager 680, by GE Healthcare (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Bicinchoninic acid assay kit, by Thermo Fisher Scientific (1 mentions)
Immobilon crescendo western hrp substrate, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Goat anti mouse igg hrp, by Enzo Life Sciences (1 mentions)
Pro prep, by iNtRON Biotechnology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Amersham imager, by Cytiva (1 mentions)
Hplc grade water, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Glutathione (gsh), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Welgene (1 mentions)
2 protocols using goat anti rabbit igg hrp
1
Western Blot Analysis of Cell Proteins
2
Investigating BITC-Induced Apoptosis Pathways
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BITC was purchased from Selleckchem (Houston, TX, USA) and dissolved in dimethyl sulfoxide (DMSO) to create a 10 mM stock solution. A glutathione (GSH) (G4705, Sigma-Aldrich Inc., Darmstadt, Germany) stock solution was prepared by dissolving GSH in HPLC-grade water (Fisher Scientific Korea Ltd., Gangnam-gu, Seoul, Korea), and this stock solution was diluted in the culture medium before treatment of cells to obtain the final working solution. RPMI-1640 medium was purchased from Welgene Inc. (Daegu, Korea), and fetal bovine serum (FBS) was purchased from Corning Inc. (Corning, NY, USA). Antibodies against cleaved Caspase-3 (c-Cas-3) (9664), Caspase-8 (9746), Caspase-9 (9508), poly (ADP-ribose) polymerase (PARP) (9532), X-linked inhibitor of apoptosis protein (XIAP) (14334), DR4 (42533) and DR5 (8074) were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP antibodies were bought from Enzo Life Sciences (Farmingdale, NY, USA). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (sc-32233), Bcl-2 (sc-492), Bax (sc-493) and Cyt c (sc-13156) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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