Cytosolic proteins of the individual human livers (30 μg in each lane) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and subsequently transferred onto nitrocellulose membranes (0.45 μm) using a Trans-Blot® Turbo™ Transfer System (Bio-Rad, USA). The membranes were blocked in 5% non-fat dry milk/TBS-Tween-20 for 2 h. For the immunodetection of CBR1, the membranes were probed overnight with primary goat polyclonal antibody to CBR1 (1:1000, ab4148; Abcam, Cambridge, UK) diluted in TBS-Tween 20 supplemented with 1% BSA, washed four times with a TBS-Tween 20 buffer, and probed with a secondary antibody (bovine anti-goat IgG-HRP, 1:3000, Santa Cruz Biotechnology, USA) for 1 h. The signal was detected using an enhanced Amersham ECL chemiluminescence kit (GE Healthcare Life Sciences, USA) according to the manufacturer’s instructions. β-Actin (mouse monoclonal, 1:3,000, Abcam) served as the loading control. The intensity of bands was evaluated using a C-DiGit™ Blot Scanner (LICOR Biotechnology, USA). Protein levels were measured in two independent experiments.
Bovine anti goat igg hrp
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Bovine anti-goat IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and quantify goat immunoglobulins (IgG) in various immunoassays and research applications.
Automatically generated - may contain errors
Lab products found in correlation
C digit blot scanner, by LI COR (2 mentions)
Trans blot turbo transfer system, by Bio-Rad (2 mentions)
Amersham ecl, by Cytiva (2 mentions)
β actin, by Abcam (1 mentions)
Amersham ecl chemiluminescence kit, by GE Healthcare (1 mentions)
Enzalutamide, by Adooq (1 mentions)
Goat anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions)
Anti aldh1, by Santa Cruz Biotechnology (1 mentions)
Anti ar, by Cell Signaling Technology (1 mentions)
Anti ar v7, by Cell Signaling Technology (1 mentions)
Anti sox2, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit igg hrp, by Beyotime (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Rabbit anti mecp2, by Cell Signaling Technology (1 mentions)
Rabbit anti vinculin, by Abcam (1 mentions)
Realtime pcr master mix, by Toyobo (1 mentions)
β actin mouse monoclonal, by Abcam (1 mentions)
Bovine anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions)
4 protocols using bovine anti goat igg hrp
1
Quantifying Hepatic CBR1 Protein Levels
2
Enzalutamide-Mediated Regulation of Stem Cell Markers
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Enzalutamide (Cat# A10562) was purchased from Adooq Bioscience, Irvine, CA, USA, and all other reagents purchased were of GLP grade. Antibodies including Anti-AR (Cat# 5153), Anti-AR-v7 (Cat# 19672), anti-POU5F1 (OCT4) (Cat# 2750S) were purchased from Cell Signaling Technologies, Beverly, MA, USA. The anti-ALDH1 (Cat# SC-166362), anti-SOX2 (Cat# SC-365823), anti-α-GAPDH (Cat# SC-47724), goat anti-mouse IgG-HRP (Cat# SC-2005), bovine anti-goat IgG-HRP (Cat# SC-2350), and goat anti-rabbit IgG-HRP (Cat# SC-2004) antibodies were purchased from Santa Cruz Biotechnology, Dallas, TX, USA.
3
Quantifying Gene and Protein Expression in Tissues
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Total RNA from hypothalamus, pituitary, testis, or TM3 cells was extracted with TRIzol (Invitrogen) and reverse transcribed (Toyobo, Cat# FSQ-301) for qRT-PCR. Real-time PCR was performed using Realtime PCR Master Mix (Toyobo, Cat# QPK-101). Relative expression of each target gene was calculated by comparison to the expression of mouse GAPDH (Primers used, see Table S1 ). For Western blot, cells were homogenized in ice-cold RIPA buffer and centrifuged at 12,000 rpm. The supernatant was resolved by 10% SDS-PAGE, electro-transferred to a PVDF membrane, and probed with the following antibodies: rabbit anti-MeCP2 (Cell Signaling Technology; Cat# 3456; 1:1 000), rabbit anti-aromatase (Abcam; Cat# ab18995; 1:500), rabbit anti-LHCGR (Proteintech; Cat# 19968-1-AP; 1:1 000), rabbit anti-Vinculin (Abcam; Cat# ab129002; 1:5 000), bovine anti-goat IgG-HRP (Santa Cruz Biotechnology; Cat# sc2352; 1:4,000), and goat anti-rabbit IgG-HRP (Beyotime; Cat# A0208; 1:1 000) antibodies. Blotting images were acquired with a Tanon-5200 imaging system. Densitometric quantification of the target protein was determined by Image J and compared with the internal control to determine the relative expression value. The internal control was Vinculin. The representative blots were selected from at least three repeated experiments.
4
Western Blot Analysis of Hepatocyte Proteins
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Microsomal or cytosolic proteins (30 µg in each lane) of hepatocytes were separated by SDS-PAGE and subsequently transferred onto nitrocellulose membranes (0.45 µm) using a Trans-Blot ® Turbo™ Transfer System (Bio-Rad, USA). The membranes were blocked in 5 % non-fat dry milk/TBS-Tween-20 for 2 h. For the immunodetection of CYP1A1, CY-P1A2, GSTA and NQO1, the membranes were probed overnight with primary antibodies [CYP1A1 -rabbit polyclonal, 1:1000 (Novus Biologicals, USA), CYP1A2 -mouse monoclonal, 1:1000 (Novus Biologicals), GSTA -goat polyclonal, 1:3000 (Abcam, UK), NQO1 -rabbit monoclonal, 1:3000 (Novus Biologicals)] diluted in TBS-Tween 20 supplemented with 1 % BSA, washed four times with TBS-Tween 20 buffer and probed with complementary secondary antibodies for 1 h [bovine anti-goat IgG-HRP, 1:3000, Santa Cruz Biotechnology (USA), bovine anti-mouse IgG-HRP, 1:10 000, Santa Cruz Biotechnology, bovine anti-rabbit IgG-HRP, 1:10 000, Santa Cruz Biotechnology]. The signal was detected using an enhanced Amersham ECL chemiluminescence kit (GE Healthcare Life Sciences, USA) according to the manufacturer's instructions. β-actin (mouse monoclonal, 1:3000, Abcam) served as the loading control. The intensity of bands was evaluated using a C-DiGit™ Blot Scanner (LI-COR Biotechnology, USA). Protein levels were measured in two independent experiments.
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