Bovine serum albumin (bsa)

Manufactured by Research Products International

Sourced in United States

The BSA is a laboratory instrument used for measuring the absorbance of solutions. It is commonly used to determine the concentration of proteins in a sample by utilizing the Beer-Lambert law. The BSA measures the amount of light absorbed by a sample at a specific wavelength, which is directly proportional to the concentration of the substance being analyzed.

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Lab products found in correlation

Triton x 100, by Thermo Fisher Scientific (3 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (3 mentions) Hel antigen, by Merck Group (2 mentions) Hrp labeled anti igg1 antibodies, by Southern Biotech (2 mentions) Ovalbumin (ova), by Merck Group (2 mentions) Spectramax m5, by Molecular Devices (2 mentions) Np1 rsa, by LGC (2 mentions) Np25 bsa, by LGC (2 mentions) Np10 bsa, by LGC (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Xylene, by Thermo Fisher Scientific (2 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions) Kynurenic acid, by Merck Group (2 mentions) Phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Irdye 800cw, by LI COR (1 mentions) Nupage lds sample buffer 4x, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Ultracomp ebeads, by Thermo Fisher Scientific (1 mentions) Tween 20, by Merck Group (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)

21 protocols using bovine serum albumin (bsa)

Cardiovascular Progenitor Cell Characterization

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The CPCs were cultured, trypsinized, and aliquoted for flow cytometry to evaluate the expression of cardiovascular progenitor cell markers including Islet-1, C-kit, CD105, and SSEA-4. The trypsinized cells were washed with 1X PBS (Life Technologies, Grand Island, NY, USA) containing 0.5% BSA (Research Products International Corp, Mount Prospect, IL, USA) and 2mM EDTA (Sigma Aldrich, St. Louis, MO, USA). The treated CPCs were fixed with 4% paraformaldehyde (PFA) (Sigma Aldrich, St. Louis, MO, USA), permeabilized in 0.1% Tween-20 (Sigma Aldrich, St. Louis, MO, USA), blocked in 0.6 M glycine (Sigma Aldrich, St. Louis, MO, USA) solution containing 10% BSA (Research Products International Corp, Mount Prospect, IL, USA), and stained for cardiovascular progenitor cell lineage markers. Once stained, the CPCs were analyzed in a MACSQuant analyzer (Miltenyi Biotec, Auburn, CA, USA). Data quantification was accomplished using FlowJo 10.7 (Ashland, OR, USA). UltraComp eBeads (Life Technologies, Grand Island, NY, USA) were used to compensate, following the manufacturer’s directions.

Knox C., Camberos V., Ceja L., Monteon A., Hughes L., Longo L, & Kearns-Jonker M. (2021). Long-Term Hypoxia Maintains a State of Dedifferentiation and Enhanced Stemness in Fetal Cardiovascular Progenitor Cells. International Journal of Molecular Sciences, 22(17), 9382.

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Quantitative Western Blot Analysis

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For protein quantification, additional five samples that were cultured in parallel were lysed in lysis buffer (RIPA buffer, catalog #89901, Thermo Fisher Scientific), phosphatase inhibitor cocktail (#78420, Thermo Fisher Scientific) and protease inhibitor cocktail (#P8340, Sigma-Aldrich), and Pierce BCA protein assay kit was used for total protein quantification (#23227, Thermo Fisher Scientific). NuPAGE LDS sample buffer 4X (#NP0007, Thermo Fisher Scientific) and β-mercaptoethanol (#6010, Calbiochem, Billerica, MA) were added to cell lysates, and samples were denaturized at 95 °C for 5 min. Proteins separation occurred on 8–12% SDS-PAGE gels and with subsequent transfer to nitrocellulose membranes. Membranes were then incubated for 1 h with blocking solution (BSA, #A30075-100, Research Products International, Mount Prospect, IL), and with primary antibodies overnight at 4 °C. Membranes were washed three times, then incubated for 1 h with secondary antibodies conjugated to infrared fluorescent dyes excitable at 800 nm (IRDye 800CW, Li-COR-USA) to visualize the desired antigens using an 800 nm laser scanner (Odyssey, Licor, USA). Quantification of protein levels was performed by IR fluorescence detection (Odyssey, Licor, USA) and analysis was performed with ImageJ.

Secchi C., Benaglio P., Mulas F., Belli M., Stupack D, & Shimasaki S. (2021). FOXO1 mitigates the SMAD3/FOXL2C134W transcriptomic effect in a model of human adult granulosa cell tumor. Journal of Translational Medicine, 19, 90.

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GHRH Antagonist Effects on Amyloid-beta and Sleep

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For acute GHRH antagonist administration, the baseline EEG and ISF Aβ was collected for 24 hr. The microdialysis perfusion buffer was artificial cerebrospinal fluid containing 0.15% BSA (Research Products International). On the experimental day, 10 μM GHRH antagonist [Phenylacetyl-(D-Arg^2.28,p-chloro-Phe⁶,Homoarg^9.29,Tyr(Me)¹⁰,Abu¹⁵,Nle²⁷)-GRF (1-29) amide; Bachem] (Varga et al., 1999 (link)) was infused at 1 μl/min in the brain via reverse-microdialysis into 3 month old PS1APP/GHRHR^+/+ males 6 hr after light onset. The EEG/EMG and ISF data were collected for another 6 hr after GHRH antagonist administration. The negative control consisted of continuous infusion of microdialysis perfusion buffer. ISF Aβ was measured with a sandwich ELISA using anti-Aβ_13-28 m266 (a gift from Eli Lilly and Company) as the coating antibody and anti-Aβ_1-5 3D6-biotin (a gift from Eli Lilly and Company) as the detecting antibody (Castellano et al., 2011 ). For data analysis, paired t-tests were performed to compare averaged ISF Aβ or sleep on the same periods of baseline day and GHRH antagonist administration day. Data were expressed as mean ±S.E.M.

Liao F., Zhang T.J., Mahan T.E., Jiang H, & Holtzman D.M. (2014). Effects of growth hormone-releasing hormone on sleep and brain interstitial fluid amyloid-β in an APP transgenic mouse model. Brain, behavior, and immunity, 47, 163-171.

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Immunocytochemical Staining of NF200

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After 24 h in culture, media was removed and washed three times with Phosphate-Buffered Saline without Calcium and Magnesium (PBS(−/−)). Cells were fixed by adding 4% paraformaldehyde in PBS(−/−) (Cat #AAJ19943K2, Fisher Scientific) for 20 min at RT. Following this, cells were washed with PBS(−/−) three times and then blocking buffer (5% normal goat serum [Cat #PCN5000, ThermoFisher], 0.2% Triton™ X-100 [Cat #9002–93-1, Fisher Scientific], and 1% BSA [Cat #9048–46-8, Research Products International] in PBS[−/−]) was added for 30 min. Chicken polyclonal antibody to NF200 (RRID: AB_2313552, Aves) was then added (1:800 in blocking buffer) and incubated at RT for 2 hours. Culture surface was washed 3 times with PBS (−/−) followed by addition of Goat anti-Chicken Alexa Fluor®546 (RRID: AB_2534097, ThermoFisher) (1:1000) for 1 hour at RT in the dark. Following this, cells were washed with PBS (−/−) three times and then cells were mounted using Fluoromount-G® (SouthernBiotech) in the dark for 24 h before imaging.

Vecchi J.T., Mullan S., Lopez J.A., Hansen M.R., Sonka M, & Lee A. (2021). NeuriteNet: A convolutional neural network for assessing morphological parameters of neurite growth. Journal of neuroscience methods, 363, 109349.

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Chemical and Reagent Acquisition

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4‐AP and kynurenic acid were obtained from Sigma‐Aldrich. KA was from Tocris. Lacosamide was from the European Pharmacopeia (through Sigma‐Aldrich). PBS, Triton X‐100, and xylenes were from Fisher Chemical. BSA was obtained from Research Products International. PFA was from Thermo Fisher Scientific.

Langton R.L., Sharma S., Tiarks G.C., Bassuk A.G, & Glykys J. (2022). Lacosamide decreases neonatal seizures without increasing apoptosis. Epilepsia, 63(12), 3051-3065.

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Dechlorionation and Staining of Embryos

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Dechorionated embryos were transferred into a scintillation vial containing a 1:1 mixture of heptane (Sigma-Aldrich) and methanol (Fischer chemical), and de-vitellinized by vigorous shaking. Once the mixture had settled, the heptane (top phase) was removed from the vial, and the embryos in methanol were transferred into a 1.5 mL microcentrifuge tube. Embryos were allowed to settle, washed once with 100% methanol, and stored at 4°C overnight (or up to 1 month). De-vitellinized embryos were rinsed with methanol and then rehydrated for 30 min in a solution of 1X PBS, 0.2% BSA (Research Products International) and 0.1% Triton X (Sigma Aldrich). Rehydrated embryos were washed with 1X PBS for 10 min, stained with DAPI (1 ug/mL in 1X PBS) (Sigma Aldrich) for 6 min, rinsed once with 1X PBS, and then mounted in Vectashield (Vector Laboratories) for imaging on a Nikon Ti2 microscope. All incubation steps for rehydration, washing, and staining were carried out at room temperature with rocking.

Kingston E.R., Blodgett L.W, & Bartel D.P. (2022). Endogenous transcripts direct microRNA degradation in Drosophila, and this targeted degradation is required for proper embryonic development. Molecular cell, 82(20), 3872-3884.e9.

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Western Blot Analysis of Protein Targets

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Cells were lysed in buffer consisting of 20 mM Tris (pH 7.5), 150 mM NaCI, 1% Triton X-100, 5 mM EDTA, 1 mM EGTA, 10μL/mL HALT Protease & Phosphatase Inhibitor Cocktail, (ThermoFisher Scientific, Grand Island, NY) on ice for 5 min. Lysates were centrifuged at 13 000 rpm a 4°C for 5 min to remove cellular debris, separated by SDS-PAGE gel electrophoresis and transferred onto a nitrocellulose membrane. (Bio-Rad, Hercules, CA). Membranes were blocked with 5% BSA (Research Products International Corp., Mt. Prospect, IL), incubated with primary antibodies against target proteins, and then incubated with horseradish peroxidase-conjugated secondary anti-bodies (Bio-Rad). The proteins were visualized using LumiGLO ECL chemiluminescence detection system (Cell Signaling Technology, Inc., Danvers, MA). The following primary antibodies were used: anti-GRB7 (Dr. Shiuh-Wen Luoh), anti-β-actin (#4967), anti-HER-1 (#4267), anti-HER-2 (#2242), anti-phospho-EGFR (#2220 and #3777), anti-β-tubulin (#3873), anti-GAPDH (#2118) (Cell Signaling Technology, Inc.).

Luoh S.W., Wagoner W., Wang X., Hu Z., Lai X., Chin K., Sears R, & Ramsey E. (2019). GRB7 dependent proliferation of basal-like, HER-2 positive human breast cancer cell lines is mediated in part by HER-1 signaling. Molecular carcinogenesis, 58(5), 699-707.

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Evaluation of Antigen-Specific IgG1 Antibodies

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Serum was harvested from blood collected by lateral tail vein sampling or cardiac puncture postmortem. OVA-specific and NP-specific IgG1 ELISA was performed as described (Brooks et al., 2020 (link); Tan et al., 2020 (link)). Briefly, ELISA plates were coated with NP conjugates (NP1-RSA, NP25-BSA, NP10-BSA all from LGC Biosearch; NP3-OVA, NP7-OVA, NP9-OVA and NP19-OVA were conjugated in-house as described above), OVA (Sigma Aldrich), BSA (Research Products International) or HEL antigen (10μg/mL; Sigma Aldrich), samples were added, and HRP-labeled anti-IgG1 antibodies (Southern Biotech) were used to detect plate-bound IgG1. Plates were developed with slow kinetic form TMB (Sigma Aldrich) and stopped with 1N sulfuric acid. Absorbance was measured at 450nm using spectrophotometer (SpectraMax M5, Molecular Devices). Relative titers were interpolated from standard curves generated using samples with known high titers of antigen-specific antibody.

Brooks J.F., Tan C., Mueller J.L., Hibiya K., Hiwa R., Vykunta V, & Zikherman J. (2021). Negative feedback by NUR77/Nr4a1 restrains B cell clonal dominance during early T-dependent immune responses. Cell reports, 36(9), 109645.

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Evaluation of Antigen-Specific IgG1 Antibodies

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Immunofluorescence Staining of Endothelial Cells

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Coverslips were coated with human fibronectin at 10 μg/ml, and cells were plated and allowed expand to confluency. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% triton and washed with PBS. Cells were blocked with 10% BSA (Sigma) in PBS. The following reagents and primary antibodies were diluted in 10% BSA (Research Products International) and allowed to incubate O/N: Phalloidin conjugated with alexa-594 was used to visualize actin 1:1000, DAPI was used to visualize the nucleus 1:1000, monoclonal VE-Cadherin antibody at 1:500 (Invitrogen; 14-1449-82), monoclonal ENOS at 1:250 (Cell Signaling; D9A5L), and monoclonal Pecam-1 at 1:500 (Cell Signaling; 89C2). Cells were washed with PBS and the following secondary antibodies were diluted 1:1000 in 10% BSA anti-rabbit alexa-488 and/or anti-mouse alexa-647 and allowed to incubate for 2 hours at room temperature. Cells were washed with PBS and mounted on coverslips using MOWIAL (Sigma Corporation).

Cronin N.M., Dawson L.W, & DeMali K.A. (2024). Mechanical activation of VE-cadherin stimulates AMPK to increase endothelial cell metabolism and vasodilation. bioRxiv.

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