Blocking one blocking buffer

Manufactured by Nacalai Tesque

Sourced in Japan

Blocking One blocking buffer is a solution used in immunoassays, such as Western blotting and ELISA, to prevent non-specific binding of antibodies. It is designed to block unoccupied binding sites on the solid support, reducing background signal and improving the specificity of the assay.

Automatically generated - may contain errors

Lab products found in correlation

Amersham imager 600, by GE Healthcare (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Phosphatase inhibitor cocktail, by Roche (1 mentions) Ecl prime western blotting detection reagent, by Cytiva (1 mentions) Phospho hsl ser563, by Cell Signaling Technology (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Wet transfer system, by Bio-Rad (1 mentions) Can get signal, by Toyobo (1 mentions) Amersham imager, by Cytiva (1 mentions) Nupage lds sample buffer, by Thermo Fisher Scientific (1 mentions) Iblot gel transfer device, by Thermo Fisher Scientific (1 mentions) Nupage novex 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Blocking One (457 citations) Blocking buffer (14 citations) Blocking One buffer (7 citations)

3 protocols using blocking one blocking buffer

Comprehensive Protein Analysis via Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole cell lysates were prepared with CHAPS buffer [40 mM HEPES (pH7.5), 120 mM NaCl, 1 mM EDTA, 0.3% CHAPS, 50 mM NaF, 1.5 mM Na₃VO₄, 10 mM glycerophosphate, 10 mM pyrophosphate, 1 mM PMSF] supplemented with protease inhibitor cocktail (Roche Diagnostics, Basel, Switzerland) and phosphatase inhibitor cocktail (Roche Diagnostics). Protein concentration was measured using the BCA assay (Thermo Scientific Pierce). Protein samples were separated by SDS-PAGE and transferred to PVDF membranes (Millipore) using the wet transfer system (Bio-Rad). Membranes were treated with BlockingOne blocking buffer (Nacalai Tesque) for 1 hr and incubated with primary antibody in Can Get Signal (Toyobo) overnight. After 3 washes in TBS-T (Tris buffered saline (pH 7.4) with 0.1% Tween 20), membranes were incubated with secondary antibodies against mouse or rabbit IgG with horseradish peroxidase in Can Get Signal for 1 hr. ECL Prime Western Blotting Detection reagent (Amersham) was used to detect immune complexes. Images were analyzed by Amersham Imager 600 (GE Healthcare Life Science). Primary antibodies recognizing the following proteins were used: Utx, phospho-HSL (Ser563), HSL, ATGL (all from Cell Signaling Technologies); beta-actin (Santa Cruz).

Ota K., Komuro A., Amano H., Kanai A., Ge K., Ueda T, & Okada H. (2019). High Fat Diet Triggers a Reduction in Body Fat Mass in Female Mice Deficient for Utx demethylase. Scientific Reports, 9, 10036.

+ Open protocol

+ Expand

Immunohistochemical Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunostaining was performed with primary antibodies in PBST with 10% Blocking One blocking buffer (Nacalai Tesque) for 2-3 nights at 4°C. After three washes of 10 min in PBS, slices were incubated in secondary antibodies from Jackson Immunoresearch at a concentration of 1:1000 in PBST containing 10% blocking buffer overnight at 4°C. Images were acquired with a Zeiss 710 LSM inverted confocal microscope using 10x, 20x, 60x oil immersion, or 100x oil immersion objectives. For presentation, some images were median filtered with a window radius of one pixel. Additionally, a subset of Z stacks underwent 3D interpolation with a resampling factor of one (3D Viewer, Image J) to permit rotated views of images. See Supplemental Experimental Procedures for additional information.

Nelson A, & Mooney R. (2016). The basal forebrain and motor cortex provide convergent yet distinct movement-related inputs to the auditory cortex. Neuron, 90(3), 635-648.

+ Open protocol

+ Expand

Western Blot Analysis of Mouse Retinal Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse eyes were enucleated and their corneas, lenses, choroids, and sclerae were removed. Mouse retinas were homogenized in RIPA lysis buffer (Cosmo Bio) and protein extracts were prepared using NuPAGE LDS Sample Buffer (Thermo Fisher Scientific). Forty micrograms of protein per lane were loaded onto NuPAGE Novex 4–12% Bis-Tris Gels (Thermo Fisher Scientific), separated, and then transferred to an iBlot Transfer Stack, Regular polyvinylidene difluoride membranes (Thermo Fisher Scientific) using the iBlot Gel Transfer Device (Thermo Fisher Scientific). Membranes were blocked with Blocking One blocking buffer (Nacalai Tesque, Kyoto, Japan) at 4°C overnight, and washed five times for 5 min in Tris-buffered saline plus 0.05% Tween 20. Membranes were then incubated with the Ab indicated for each experiment in blocking buffer at 4°C overnight. After washing with Tris-buffered saline plus 0.05% Tween 20, membranes were incubated with goat anti-rabbit IgG H&L (HRP) Ab (1:1,000; Abcam) in blocking buffer at 4°C overnight. Protein bands were detected using a Chemi-Lumi One L assay kit (Nacalai Tesque).

Kawashima R., Matsushita K., Mandai K., Sugita Y., Maruo T., Mizutani K., Midoh Y., Oguchi A., Murakawa Y., Kuniyoshi K., Sato R., Furukawa T., Nishida K, & Takai Y. (2024). Necl-1/CADM3 regulates cone synapse formation in the mouse retina. iScience, 27(4), 109577.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!