Bovine serum albumin (bsa)

Manufactured by Jackson ImmunoResearch

Sourced in United States, United Kingdom, China, Germany

BSA (Bovine Serum Albumin) is a common laboratory reagent used as a protein standard and stabilizer. It is derived from bovine serum and is widely used in various biochemical and immunological applications.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (13 mentions) Triton x 100, by Merck Group (10 mentions) Normal donkey serum (nds), by Jackson ImmunoResearch (7 mentions) Normal goat serum (ngs), by Jackson ImmunoResearch (4 mentions) Tween 20, by Merck Group (4 mentions) Triton x 100, by Jackson ImmunoResearch (3 mentions) Ripa buffer, by Thermo Fisher Scientific (3 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Trypsin edta, by Thermo Fisher Scientific (2 mentions) Anti ki67 d3b5 rabbit antibody, by Cell Signaling Technology (2 mentions) Dako pen, by Agilent Technologies (2 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Fluoromount, by Merck Group (2 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions) Rabbit anti iba1, by Fujifilm (2 mentions) Alexa fluor 488 conjugated goat anti chicken igg, by Thermo Fisher Scientific (2 mentions) Goat serum tbst, by Jackson ImmunoResearch (2 mentions) Alexa fluor 568 conjugated goat anti mouse igg2a, by Thermo Fisher Scientific (2 mentions)

109 protocols using bovine serum albumin (bsa)

Immunofluorescence Analysis of Phosphorylated STAT3

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Cultured LNCaP and DU-145 cells were grown on coverslips, and LNCaP were treated with IL-6 or tBHP in the same aforementioned conditions. After the stimulation, cells were washed with PBS, fixed with 4% formaldehyde for 15 min and then rinsed with PBS. Cell were then permeabilized with cold MeOH (−20 °C) for 5 min. After washing three times with PBS, cells were blocked overnight with 3% w/v BSA (Sigma-Aldrich) in PBS. Fixed cells were processed for immunofluorescence staining to detect the localization of pS⁷²⁷-STAT3 (Sigma-Aldrich, SAB4300034, antibody dilution 1:200) using a specific primary antibody diluted in PBS containing 2% w/v BSA for 1 h. Following three washes with PBS added with 0.05% Triton and 2% w/v BSA (PBS-T), cells were incubated for 1 h in the darkness with a Fluorescein isothiocyanate FITC-conjugated secondary antibody (Jackson Immunoresearch, dilution 1:800). Cell nuclei were counterstained with 100 ng/mL Hoechst 33258 for 15 min. After washing with PBS-T, coverslips were mounted on glass microscopy slides with Duolink Mounting Medium and examined with a confocal fluorescence microscope (ZEISS LSM510META) with magnification using 60x oil immersion objective lens. Images were acquired and analysed by ZEN 215 software (ZEISS, Germany).

Cocchiola R., Rubini E., Altieri F., Chichiarelli S., Paglia G., Romaniello D., Carissimi S., Giorgi A., Giamogante F., Macone A., Perugia G., Gurtner A, & Eufemi M. (2019). STAT3 Post-Translational Modifications Drive Cellular Signaling Pathways in Prostate Cancer Cells. International Journal of Molecular Sciences, 20(8), 1815.

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Immunofluorescent Staining of Alveolar Cells

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We further performed immunofluorescent staining (Novus Biologicals, USA) for alveolar type I cells. The sections were treated with 3% hydrogen peroxide, and nonspecific binding sites were blocked with bovine serum albumin (BIONOVAS, Toronto, Canada) for 30 mins. The sections were incubated with antibodies against rat type I cell 40-kDa protein (RTI40)/type I cell alpha (T1α) protein (dilution 1:50; Sino Biological, USA) and NKX2.1/type II (NKX2.2) (dilution 1:100; NOVUS Biological, Toronto, Canada) overnight at 4°C. The fluorescent label-conjugated FITC- and Texas Red-labeled secondary antibody (Jackson Immunoresearch Laboratories) was diluted with 1% bovine serum albumin in phosphate buffered saline at a ratio of 1:250 at room temperature for 1 h, respectively. We evaluated the sections using an EVOS M7000 Imaging System (Thermo Fisher Scientific, USA).

Hsieh P.C., Kuo C.Y., Wu C.P., Yue C.T., Peng C.K., Huang K.L, & Lan C.C. (2021). Nonionic surfactant attenuates acute lung injury by restoring epithelial integrity and alveolar fluid clearance. International Journal of Medical Sciences, 18(6), 1363-1374.

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FITC-DRP Uptake and Evaluation

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Cells plated at 6.4 × 10⁵ cells per well in 6-well plates were treated a day later with 3 µM doses of FITC-DRPs for 1 h or 24 h at 37 °C, 5% CO₂. Cells were washed using DPBS with calcium and magnesium (Thermo-Fisher Scientific, Cambridge, MA, USA) and dissociated from plates using 0.25% trypsin EDTA (Thermo-Fisher Scientific, Cambridge, MA, USA) at 37 °C, 5% CO₂ for 2–3 min. Dissociated cells were collected in tubes and washed twice by being spun at 1200 rpm for 5 min and resuspended in a buffer of DPBS supplemented with 1% bovine serum albumin (Jackson Immunoresearch Laboratories, West Grove, PA, USA) and 0.1% sodium azide 0.1 M solution (Millipore-Sigma, Burlington, MA, USA). Following washes, cells were run through 40 µM cell strainers (Thermo-Fisher Scientific, Cambridge, MA, USA) and loaded 250 µL/well into a clear round-bottom 96-well plate. Flow cytometry was conducted using a Guava easyCyte HT flow cytometer and guavaSoft 3.2 software (Luminex Corporation, Northbrook, IL, USA). Experiments were repeated once (executed a total of two times).

Gill A.L., Wang M.Z., Levine B., Premasiri A, & Vieira F.G. (2019). Primary Neurons and Differentiated NSC-34 Cells Are More Susceptible to Arginine-Rich ALS Dipeptide Repeat Protein-Associated Toxicity than Non-Differentiated NSC-34 and CHO Cells. International Journal of Molecular Sciences, 20(24), 6238.

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Immunofluorescent Localization of AhR

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Keratinocytes were grown on glass slides in 24-well plates. After 20 min exposure to S. mitis (MOI 1:60) the cells were washed twice in PBS, fixed and permeabilized at −25 °C for 5 min using ice cold methanol:acetone solution (7:3). For blocking, 1% IgG-free bovine serum albumin (Jackson Immunoresearch, Newmarket, UK) in PBS was added and left for 30 min at RT before incubated with polyclonal AhR goat anti-human IgG antibody (Santa Cruz, Dallas, TX, USA, 1:150) for 2 h at 4 °C. After washing twice, Cy2-conjugated donkey anti-goat IgG polyclonal secondary antibody (Jackson, 1:150) was added to each well and left for incubation for 1 h before DAPI was added and the slides were mounted on cover glasses.

Engen S.A., Rørvik G.H., Schreurs O., Blix I.J, & Schenck K. (2017). The oral commensal Streptococcus mitis activates the aryl hydrocarbon receptor in human oral epithelial cells. International Journal of Oral Science, 9(3), 145-150.

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Immunohistochemical Staining of Ki67 in Tissue Sections

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Following cryosection, slides were initially outlined with a Dako Pen (Dako, Carpinteria, CA, USA). Slides were washed 3 times in PBS, 10 minutes per wash for a total of 30 minutes and then permeabilized with 0.5% Triton (Thermo-Fisher Scientific) for 10 minutes. Slides were returned to a 10 minutes PBS wash before antigen retrieval. For antigen retrieval, slides were placed in two, 7 minute rounds of boiling PBS for a total of 14 minutes. Following retrieval, slides were washed in fresh PBS. Slides were blocked for one hour in 3% or 5% bovine serum albumin in PBS (Jackson ImmunoResearch, West Grove, PA, USA) and were then stained with anti-Ki67 (D3B5) rabbit antibody at a 1/100 concentration (Cell Signaling, Danvers, MA, USA; 91295) overnight at 4°C. The following day, slides were washed in three, 30 minute rounds of PBS for a total of 90 minutes. Slides were permeabilized with 0.1% Triton for 10 minutes before being switched into secondary antibody (Life Technologies, Carlsbad, CA, USA) Alexa-Fluor 594 goat anti-rabbit IgG at 1:300 concentration for 2 hours at room temperature. Finally, slides were washed with 3 rounds of PBS, 20 minutes each round, for a total of 60 minutes and then mounted in Fluoromount with 4′, 6-diamidino-2-phenylindole dihydrochloride (DAPI; Sigma-Aldrich) and coverslipped (Thermo-Fisher Scientific).

Welker A.M., Jaros B.D., An M, & Beattie C.E. (2017). Changes in tumor cell heterogeneity after chemotherapy treatment in a xenograft model of glioblastoma. Neuroscience, 356, 35-43.

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Cell Culture for Chromatin Tracing

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Cells were prepared similarly to our previous studies (Bintu et al., 2018 (link); Wang et al., 2016 (link)). IMR-90 cells were purchased
from American Type Culture Collection (ATCC, CCL-186) and grown in the
recommended growth media (Eagle’s Minimum Essential Media (EMEM; ATCC
30–2003), containing 10% w/v Bovine Serum Albumin (BSA; Jackson
Immunoresearch 001–000-162) and 1% v/v Penicillin-Streptomycin (Sigma
P0781)) at 37°C and 5% CO₂. Media was exchanged every 2
days and the cells were passaged when-ever the culture reached confluency
(every 2–5 days). To avoid potential alterations to chromatin
structure, all chromatin tracing experiments in this study were performed on
cells cultured for 6 weeks at most.

Su J.H., Zheng P., Kinrot S.S., Bintu B, & Zhuang X. (2020). Genome-Scale Imaging of the 3D Organization and Transcriptional Activity of Chromatin. Cell, 182(6), 1641-1659.e26.

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Reelin Immunostaining and Counterstaining Protocol

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Slices were blocked 2 h at room temperature in PBS 0.01 M/7% (v/v) normal donkey serum (017-000-121, Jackson Immuno Research)/0.3% (v/v) triton (Triton X-100, Sigma-Aldrich) and incubated overnight at 4°C with a goat Reelin antibody (AF3820, R&D Systems, 1:500) diluted in PBS 0.01 M/2% bovine serum albumin (001-000-161, Jackson Immunoresearch)/0.05% sodium azide (S002, Sigma-Aldrich)/0.1% Triton. The next day, slices were washed 3 times with PBS 0.01 M, incubated for 2 h at room temperature with 4′,6-diamidino-2-phenylindole (DAPI) and Alexa Fluor 647-coupled secondary antibody (Dianova, Hamburg, Germany) diluted in PBS 0.01 M/2% bovine serum albumin/0.05% azide, washed 3 times with PBS 0.01 M and mounted in fluoromount (Sigma-Aldrich).

Engeroff K., Warm D., Bittner S, & Blanquie O. (2023). Different activity patterns control various stages of Reelin synthesis in the developing neocortex. Cerebral Cortex (New York, NY), 33(15), 9376-9386.

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Measuring CAR Expression on T Cells

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To measure CAR expression, 0.25 million cells were suspended in 100 µL of buffer (Phosphate-buffered saline (PBS) containing 2 mM Ethylenediaminetetraacetic acid (EDTA) pH 8 and 0.5% Bovine serum albumin (BSA)) and incubated on ice with 1 µL of human serum (Jackson Immunoresearch, West Grove, PA, USA) for 10 min. Then, 0.3 µg of biotinylated human DCLK1 protein (AcroBiosystems, Newark, DE, USA) was added and the cells were incubated on ice for 30 min. Cells were rinsed with 3 mL of buffer and suspended in 100 µL of buffer. Next, 1 µL of phycoerythrin (PE)-conjugated streptavidin (BD Biosciences, San Jose, CA, USA), 1 µL of allophycocyanin (APC)-labeled anti-CD3 (BioLegend, San Diego, CA, USA) and 2 µL of 7-aminoactinomycin D (7-AAD) solution (BioLegend) were added to the cells and incubated on ice for 30 min [39 (link),42 (link)]. The cells were rinsed with 3 mL of buffer and suspended in buffer and acquired on a FACSCalibur (BD Biosciences). Cells were analyzed first for light scatter versus 7-AAD staining, and the 7-AAD⁻ live gated cells were plotted for anti-CD3 staining versus DCLK1 protein staining [39 (link),42 (link)].

Sureban S.M., Berahovich R., Zhou H., Xu S., Wu L., Ding K., May R., Qu D., Bannerman-Menson E., Golubovskaya V, & Houchen C.W. (2019). DCLK1 Monoclonal Antibody-Based CAR-T Cells as a Novel Treatment Strategy against Human Colorectal Cancers. Cancers, 12(1), 54.

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Immunocytochemistry of Aortic Smooth Muscle Cells

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Cultured primary human aortic smooth muscle cells were grown on 10 μg/ml fibronectin (Sigma F2006)‐coated glass‐bottomed dishes (MatTek CORPORATION P35G‐1.5‐20‐C). Cells were first fixed with 2% paraformaldehyde (Polysciences, Inc, 18814) in PBS for 20 min at 37°C, then permeabilized with 0.1% Triton X‐100 in PBS containing 2% PFA at room temperature for 5 min, and blocked with 3% bovine serum albumin (Jackson ImmunoResearch Laboratories, Inc. 001‐000‐162) at room temperature for 60 min. Cells were washed with PBS and incubated with SM α‐actin‐Cy3 (1:1,000 in 1% BSA), SM22α (1:1,000 in 1% BSA), and SM‐calponin (1:500 in 1% BSA) antibodies at 4°C overnight, washed three times with PBS, and incubated with diluted Alexa Fluor‐conjugated secondary antibody (1:500) (Life Technologies) for 1 h at room temperature. The dishes were then washed three times with PBS and mounted using Prolong Gold antifade reagent with DAPI (Life Technologies P36931).

Chen P., Qin L., Li G., Tellides G, & Simons M. (2016). Smooth muscle FGF/TGFβ cross talk regulates atherosclerosis progression. EMBO Molecular Medicine, 8(7), 712-728.

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Lectin-Based HbA1c Quantification

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All biotinylated lectins described in this report were obtained from J-oil Mills, Inc., Tokyo, Japan. ELISA plates (half area 96 well, flat bottom) were purchased from Greiner Bio-One, Frickenhausen, Germany. Human hemoglobin was obtained from Sigma-Aldrich (St. Louis, MO, USA). Highly purified HbA_1c was purchased from BBI Solutions (Cardiff, UK). Monoclonal antibody against HbA_1c was purchased from Abnova Corp. (Taipei, Taiwan). Bovine serum albumin for the blocking of the plates was purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). All the other reagents were purchased from Wako Pure Chemical Industries (Osaka, Japan), unless otherwise stated.

Kabata M., Hase E., Kimura K., Kobayashi Y., Ueno Y, & Yoshimune K. (2016). Assay of hemoglobin A1c using lectin from Aleuria aurantia. AMB Express, 6, 119.

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