Ab181440

Manufactured by Abcam

Sourced in United Kingdom

Ab181440 is a primary antibody that recognizes the androgen receptor (AR) protein. It is designed for use in various research applications, including Western blotting, immunohistochemistry, and immunofluorescence.

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Lab products found in correlation

Chemidoc xrs imaging system, by Bio-Rad (1 mentions) Protein assay reagent, by Thermo Fisher Scientific (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Mini protean tgx gel, by Bio-Rad (1 mentions) Mouse monoclonal antibody to β actin, by Merck Group (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Sp 9001 splink detection kits, by OriGene (1 mentions) Il 18 antibody, by Proteintech (1 mentions) Mmp 2 antibody, by Proteintech (1 mentions) T6074, by Merck Group (1 mentions) Ab21286, by Abcam (1 mentions) Ab108337, by Abcam (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Nupage western blot system, by Thermo Fisher Scientific (1 mentions)

4 protocols using ab181440

Western Blot Analysis of Osteopontin

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Cells were plated in 6-well culture plates the day before treatment/infection and harvested in lysis buffer containing 1% NP-40, 150 mM NaCl, 50 mM Tris-HCl pH 8.0 and a protease inhibitor cocktail (Thermo Fisher Scientific). After removal of cellular debris by centrifugation, total protein concentration was measured at 660nm using protein assay reagent (Thermo Fisher Scientific) and 25 μg of total protein were separated in a precast 12% mini-PROTEAN TGX gel (Bio-Rad) and transferred to a nitrocellulose membrane (Bio-Rad). The membrane was incubated with a rabbit polyclonal antibody to human OPN (Abcam, ab181440) and a mouse monoclonal antibody to β-actin (Sigma-Aldrich); proteins were detected by incubating with a secondary anti-mouse IgG-HRP and/or anti-rabbit IgG-HRP, followed by the ECL reagent kit (Pierce). Images were captured using a ChemiDoc XRS+ imaging system (Bio-Rad).

Sampayo-Escobar V., Green R., Cheung M.B., Bedi R., Mohapatra S, & Mohapatra S.S. (2018). Osteopontin plays a pivotal role in increasing severity of respiratory syncytial virus infection. PLoS ONE, 13(4), e0192709.

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Western Blot Analysis of Osteogenic Markers

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Total cellular protein was isolated from the cultured cells using RIPA extraction buffer and the concentration was determined by BCA assay. A total of 30 µg protein per well was separated by 10–15% SDS-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes, which were blocked for 2 h at 24–25°C with 5% BSA/TBST and then incubated overnight at 4°C with the following primary antibodies: rabbit monoclonal anti-BMP2 (1:1,000; ab183729), mouse monoclonal anti-osterix (1:1,000; ab57335), rabbit polyclonal anti-OPN (1:1,000; ab181440) and rabbit monoclonal anti-ALP (1:1,000; ab186422) (all from Abcam, Cambridge, UK); and mouse monoclonal anti-GAPDH (1:2,000; BE0023; Bioeasytech, Beijing, China). Immunoreactivity was detected with IRDye 800cw-conjugated goat anti-rabbit/-mouse IgG (1:10,000; LI-COR Biosciences, Lincoln, NE, USA) secondary antibodies and visualized with an Odyssey infrared imaging system (LI-COR Biosciences), with gray values analyzed using Odyssey v3.0 software.

Ning S., Chen Z., Fan D., Sun C., Zhang C., Zeng Y., Li W., Hou X., Qu X., Ma Y, & Yu H. (2016). Genetic differences in osteogenic differentiation potency in the thoracic ossification of the ligamentum flavum under cyclic mechanical stress. International Journal of Molecular Medicine, 39(1), 135-143.

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Immunohistochemical Analysis of Inflammatory Markers

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Paraformaldehyde-fixed paraffin-embedded sections were incubated with primary antibodies overnight at 4°C. The primary antibodies used were against OPN-N (rabbit anti-OPN-N antibody, ab181440, 1:200; Abcam, United Kingdom), NLRP3 (rabbit anti-NLRP3 antibody, 1:200; Boster, Wuhan, China), ASC (rabbit anti-ASC antibody, 10500-1-AP, 1:200; Proteintech, Wuhan, China), IL-18 (rabbit anti- IL-18 antibody, 1:200; Proteintech), IL-1β (rabbit anti-IL-1β antibody, 1:200; Arigo, Hamburg, Germany), Caspase-1 (rabbit anti- Caspase-1 antibody, 22915-1-AP, 1:200; Proteintech), MMP2 (rabbit anti-MMP2 antibody, 1:200; Proteintech), MMP9 (rabbit anti-MMP9 antibody, 1:200; Proteintech), α-smooth muscle actin (SMA) (rabbit anti-α-SMA, 1:200; Proteintech) and α9β1 (rabbit anti-α9β1, ab27947, 1:200; Abcam). As negative controls, species and isotype-matched IgG was applied in place of the primary antibody. Immunohistochemical analysis was performed using the SP-9001 SPlink Detection kits (OriGene Technologies, Inc.), according to the manufacturers’ instructions. All sections were examined using a Nikon E100 microscope. Three sections were randomly selected from each group.

Liu H., Zhang Y., Song W., Sun Y, & Jiang Y. (2021). Osteopontin N-Terminal Function in an Abdominal Aortic Aneurysm From Apolipoprotein E-Deficient Mice. Frontiers in Cell and Developmental Biology, 9, 681790.

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Western Blot Analysis of Bone Proteins

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Freshly dissected bones of WT and Smpd3 -/-male and female mice were homogenized mechanically in lysate buffer containing protease inhibitor cocktail (Complete; Roche, Penzberg, Germany). Protein concentrations were measured using the Pierce BCA protein assay kit (Thermo Fisher Scientific, Darmstadt, Germany). Protein aliquots (100 mg) were separated by NuPAGE 4% to 12% BIS-TRIS gels and transferred to a nitrocellulose membrane, using the NuPAGE Western Blot system (Invitrogen, Darmstadt, Germany). Blots were immunostained overnight at 4 C with the following respective antibodies: antiealkaline phosphatase (1:2000, ab108337, RRID:AB_10862036), antiecollagen I (1:500, ab21286, RRID:AB_446161), and anti-osteopontin (1:1000, ab181440) (all from Abcam, Cambridge, UK), and antiea tubulin (1:12,000, T6074, RRID:AB_477582; Sigma-Aldrich, St. Louis, MO). After washing, horseradish peroxidaseeconjugated secondary antibodies were used and detected with the enhanced chemiluminescence system. Signals were quantified by densitometry using the ImageJ2 version 2.0.0-rc-3 (NIH, Bethesda, MD) program (RRID:SCR_003070).

Stoffel W., Hammels I., Jenke B., Schmidt-Soltau I, & Niehoff A. (2019). Neutral Sphingomyelinase 2 (SMPD3) Deficiency in Mice Causes Chondrodysplasia with Unimpaired Skeletal Mineralization. The American journal of pathology, 189(9).

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