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casein based blocking solution  (Thermo Fisher)


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    Thermo Fisher casein based blocking solution
    Casein Based Blocking Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/casein+based+pbs+blocking+buffer/Casein+Blocking+Buffer%2C+3%25+in+PBS/pmc12522083-92-32-35
    Average 95 stars, based on 1 article reviews
    casein based blocking solution - by Bioz Stars, 2026-11
    95/100 stars

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    Incubation:

    Article Title: Generation of human sequence antibodies in OmniChicken against the highly conserved mammalian target BDNF using three distinct discovery workflows.
    Article Snippet: .. Following overnight incubation, periplasmic extract was prepared in Tris-EDTA-sucrose buffer (TES; 50 mM Tris-HCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 20% Sucrose, pH 8) and blocked with 1% Casein/PBS (Thermo Scientific) to obtain a final concentration of 50% blocked periplasmic extract. .. Ninety-six well NUNC Maxisorp plates were pre-coated overnight at 4°C with 100 μL BDNF (Sino Biologics Inc, 50,240) at a concentration of 5 μg/mL in PBS pH 7.4.

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Article Title: ATM functions as a rheostat of metabolic stress in small-cell lung cancer
    Article Snippet: Equal amounts of protein (50 μg) were mixed with Laemmli Sample Buffer (Bio-Rad, 1610747), separated on 10-15% SDS-PAGE gels, and transferred by wet blotting onto 0.45-μm Immobilon-FL PVDF membranes (Millipore, IPFL00010). .. Membranes were blocked for 1 h at room temperature in Blocker Casein (Thermo Scientific, 37532) prepared in TBS containing 0.1% Tween 20 (Fisher Bioreagents, BP337-500), then incubated overnight at 4 °C with primary antibodies (1:1000 dilution in 5% bovine serum albumin [BSA]). ..

    Concentration Assay:

    Article Title: Generation of human sequence antibodies in OmniChicken against the highly conserved mammalian target BDNF using three distinct discovery workflows.
    Article Snippet: .. Following overnight incubation, periplasmic extract was prepared in Tris-EDTA-sucrose buffer (TES; 50 mM Tris-HCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 20% Sucrose, pH 8) and blocked with 1% Casein/PBS (Thermo Scientific) to obtain a final concentration of 50% blocked periplasmic extract. .. Ninety-six well NUNC Maxisorp plates were pre-coated overnight at 4°C with 100 μL BDNF (Sino Biologics Inc, 50,240) at a concentration of 5 μg/mL in PBS pH 7.4.

    Radio Immunoprecipitation:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Centrifugation:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Protein Concentration:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Acid Assay:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Bicinchoninic Acid Protein Assay:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Membrane:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Saline:

    Article Title: Exertional heat stroke-related acute kidney injury: novel animal and cellular models
    Article Snippet: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, dojindo, CK04, 1:10) according to the manufacturer’s instructions. .. after the cells were lysed using radio Immunoprecipitation assay (rIPa, Servicebio, G2002, 1:2) buffer containing phenylmethylsulfonyl fluoride (PMSF, Solarbio, P0100, 1:100), the supernatant was collected by centrifugation, and the protein concentration was assessed using a bicinchoninic acid assay (bCa) protein assay kit (thermo, 23227). a total of 20 μg of protein was loaded per lane onto a 12% sodium dodecyl sulfate-polyacrylamide gel (SdS-PaGE, bIo-rad, 4561043) and subsequently transferred to a nitrocellulose (NC) membrane. the membrane was blocked at room temperature with a 1× casein solution (50 μL, thermo, 37582) for 2 h and then incubated overnight at 4 °C with the following antibodies: neutrophil gelatinase-associated lipocalin (NGaL, 10 mL, abcam, ab26426, 1:1000) and β-tubulin (10 mL, Proteintech, 66240-1, 1:20, 000). the membrane was then incubated with the corresponding secondary antibody (10 mL, beyotime, a0208, 1:2000) at room temperature for 2 h, followed by three washes with tris buffered saline with tween (tbSt, Solarbio, t1081, 1:10). .. Signals were detected using the ChemidocTM touch Imaging System (bio-rad, Hercules, Ca, uSa) with enhanced chemiluminescence (ECL, 2 mL, amersham biosciences, GErPN2235, 1:1), and the data were analyzed using ImageJ software (version 1.8.0, Md, uSa).

    Blocking Assay:

    Article Title: Analysis of GRK2 aggregation in the pathology of Alzheimer disease in animal models
    Article Snippet: Co-localization of TOMM40 and GRK2 on hippocampal specimens was performed with affinity-purified, rabbit polyclonal anti-TOMM40 antibodies (18409-1-AP; Proteintech) and mouse monoclonal anti-GRK2 antibody (sc-13143; Santa Cruz Biotechnology Inc.). .. Primary antibodies in blocking buffer (PBS with 5% [w/v] bovine serum albumin fraction V, 0.05% Tween 20, pH 7.4) were applied at a dilution of 1:400 for 1 h. After three washing steps with PBS supplemented with 0.05% Tween 20 to remove unbound antibodies, secondary antibodies labeled with Alexa Fluor 488 (Alexa Fluor 488 goat anti-Mouse IgG [H + L] Highly Cross-Adsorbed Secondary Antibody; A-11029; Invitrogen Molecular Probes) and labeled with Alexa Fluor 568 (Alexa Fluor 568 goat anti-Rabbit IgG (H + L); A-11011; Invitrogen by Thermo Fisher Scientific) were applied in blocking buffer at a dilution of 1:2000. .. After blocking of the PVDF membrane with PBS supplemented with 0.05% (v/v) Tween 20 and 5% bovine serum albumin or non-fat dried milk for 1 h, primary antibodies were applied at a dilution of 1:2000 - 1:5000 in blocking buffer for 1 h. Unbound antibodies were removed by three washing steps with PBS supplemented with 0.05% (v/v) Tween 20, and the bound antibodies were visualized with F(ab) 2 fragments of enzyme-coupled secondary antibodies (Jackson ImmunoResearch Laboratories; dilution 1:40,000), by peroxidase-coupled protein A (Calbiochem, Merck Millipore; dilution 1:10,000), or by peroxidase-coupled protein G (EMD Millipore Corporation; dilution 1:10,000), as applicable, followed by enhanced chemiluminescent detection (ECL Prime, GE Healthcare Cytiva; ECL Plus, Thermo Fisher Scientific).

    Labeling:

    Article Title: Analysis of GRK2 aggregation in the pathology of Alzheimer disease in animal models
    Article Snippet: Co-localization of TOMM40 and GRK2 on hippocampal specimens was performed with affinity-purified, rabbit polyclonal anti-TOMM40 antibodies (18409-1-AP; Proteintech) and mouse monoclonal anti-GRK2 antibody (sc-13143; Santa Cruz Biotechnology Inc.). .. Primary antibodies in blocking buffer (PBS with 5% [w/v] bovine serum albumin fraction V, 0.05% Tween 20, pH 7.4) were applied at a dilution of 1:400 for 1 h. After three washing steps with PBS supplemented with 0.05% Tween 20 to remove unbound antibodies, secondary antibodies labeled with Alexa Fluor 488 (Alexa Fluor 488 goat anti-Mouse IgG [H + L] Highly Cross-Adsorbed Secondary Antibody; A-11029; Invitrogen Molecular Probes) and labeled with Alexa Fluor 568 (Alexa Fluor 568 goat anti-Rabbit IgG (H + L); A-11011; Invitrogen by Thermo Fisher Scientific) were applied in blocking buffer at a dilution of 1:2000. .. After blocking of the PVDF membrane with PBS supplemented with 0.05% (v/v) Tween 20 and 5% bovine serum albumin or non-fat dried milk for 1 h, primary antibodies were applied at a dilution of 1:2000 - 1:5000 in blocking buffer for 1 h. Unbound antibodies were removed by three washing steps with PBS supplemented with 0.05% (v/v) Tween 20, and the bound antibodies were visualized with F(ab) 2 fragments of enzyme-coupled secondary antibodies (Jackson ImmunoResearch Laboratories; dilution 1:40,000), by peroxidase-coupled protein A (Calbiochem, Merck Millipore; dilution 1:10,000), or by peroxidase-coupled protein G (EMD Millipore Corporation; dilution 1:10,000), as applicable, followed by enhanced chemiluminescent detection (ECL Prime, GE Healthcare Cytiva; ECL Plus, Thermo Fisher Scientific).



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