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Bovine serum albumin (bsa)

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Bovine serum albumin (BSA) is a protein derived from bovine serum. It is commonly used as a reagent in various laboratory applications due to its ability to stabilize and protect other proteins. BSA serves as a blocking agent, stabilizer, and carrier protein in numerous biochemical assays and experiments.

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Lab products found in correlation

Kaad cyclopamine, by Merck Group (2 mentions) Recombinant shh c25ii, by R&D Systems (2 mentions) Ihh c28ii, by GenScript (2 mentions) N acetyl l cysteine, by Merck Group (2 mentions) Toxinsensor gel clot endotoxin assay kit, by GenScript (1 mentions) Glutaraldehyde, by Sinopharm (1 mentions)

4 protocols using bovine serum albumin (bsa)

1

Expansion and Preparation of Antibodies

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5E1 antibody was expanded in our laboratory (see supplemental material and methods) and prepared in PBS. IgG1 (InVivoMab, BE0083) was diluted in PBS. KAAD-cyclopamine (Millipore) was prepared in DMSO. Recombinant SHH (C25II) (R&D Systems) and IHH (C28II) (Genscript) were prepared in PBS containing 0.1% bovine serum albumin (BSA). N-Acetyl-L-cysteine (NAC) was purchased from Sigma-Aldrich and prepared in PBS for i.p. injection or sterile tap water for supplemented drinking water. For NAC solution, pH was adjusted to 7.4.
Kasiri S., Chen B., Wilson A.N., Reczek A., Mazambani S., Gadhvi J., Noel E., Marriam U., Mino B., Lu W., Girard L., Solis L.M., Luby-Phelps K., Bishop J., Kim J.W, & Kim J. (2020). Stromal Hedgehog pathway activation by IHH suppresses lung adenocarcinoma growth and metastasis by limiting reactive oxygen species. Oncogene, 39(16), 3258-3275.
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2

Preparation of Reagents for SHH/IHH Study

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5E1 antibody was expanded in our laboratory (see Supplementary material and methods) and prepared in PBS. IgG1 (InVivoMab, BE0083) was diluted in PBS. KAAD-cyclopamine (Millipore) was prepared in DMSO. Recombinant SHH (C25II) (R&D Systems) and IHH (C28II) (Genscript) were prepared in PBS containing 0.1% bovine serum albumin (BSA). N-Acetyl-L-cysteine (NAC) was purchased from Sigma-Aldrich and prepared in PBS for i.p. injection or sterile tap water for supplemented drinking water. For NAC solution, pH was adjusted to 7.4.
Kasiri S., Chen B., Wilson A.N., Reczek A., Mazambani S., Gadhvi J., Noel E., Marriam U., Mino B., Lu W., Girard L., Solis L.M., Luby-Phelps K., Bishop J., Kim J.W, & Kim J. (2020). Stromal Hedgehog pathway activation by IHH suppresses lung adenocarcinoma growth and metastasis by limiting reactive oxygen species. Oncogene, 39(16), 3258-3275.
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3

Purification of rAceyCP1, rAceyCPL, and rAceySKPI3

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rAceyCP1, rAceyCPL and rAceySKPI3 were purified from X-33 culture supernatants by immobilized metal affinity chromatography using a Ni resin and column (GenScript). Proteins bound to the resin were washed with Triton X-100 to reduce endotoxin levels to <1 EU/μg. The eluates were buffer exchanged into PBS (pH 7.4) by dialysis, and then filter sterilized with 0.22 μm Millex-GP Syringe Filters. Endotoxin levels were detected by ToxinSensor Gel Clot Endotoxin Assay Kit (GenScript). Protein concentrations were determined by Bradford assay using BSA as standard (GenScript).
Noon J.B., Schwarz E.M., Ostroff G.R, & Aroian R.V. (2019). A highly expressed intestinal cysteine protease of Ancylostoma ceylanicum protects vaccinated hamsters from hookworm infection. PLoS Neglected Tropical Diseases, 13(4), e0007345.
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4

Peptide-BSA Conjugation Protocol

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Chemical conjugation of untagged STh mutant peptides to BSA was performed by GenScript (Leiden, The Netherlands). BSA (Amresco 0332, Quality Biological Inc., Gaithersburg, MD, USA) was dissolved in PBS to a concentration of 10 mg/mL and mixed with selected STh mutant peptides (STh-A14T, STh-L9A/A14T, STh-L9K/A14T or STh-L9N/A14T) at 1:1 mass ratios, followed by dilution with PBS to half the final reaction volume (1 mg/mL). Next, 0.25% glutaraldehyde (Sinopharm Chemical Reagent Co., Ltd. cat # 30092436, Shanghai, China) (half final volume) was slowly added and placed in a constant temperature oscillator at 25 ± 2 °C for 2 h. To quench unreacted glutaraldehyde, 1% glycine was subsequently added and further incubated at 25 ± 2 °C for 1 h in dark. To remove STh mutant peptides not covalently linked to BSA, the conjugate samples were dialyzed against PBS using a 14 kDa MWCO dialysis membrane at room temperature for 2 h, followed by a change of fresh PBS buffer and overnight (~15 h) at 4 °C.
Zegeye E.D., Diaz Y, & Puntervoll P. (2022). Vaccine Candidate Double Mutant Variants of Enterotoxigenic Escherichia coli Heat-Stable Toxin. Vaccines, 10(2), 241.
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