Low endotoxin bovine serum albumin bsa

Manufactured by Merck Group

Sourced in United States

Low endotoxin bovine serum albumin (BSA) is a purified protein derived from bovine serum. It is designed to have low levels of endotoxins, which are bacterial components that can potentially interfere with cell culture and other experimental applications.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (2 mentions) Ultrapure lps, by InvivoGen (1 mentions) Raw 264.7 cells, by American Type Culture Collection (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Penicillin, by MP Biomedicals (1 mentions) Streptomycin, by MP Biomedicals (1 mentions) Anti tlr4 sc 293072, by Santa Cruz Biotechnology (1 mentions) Primescript rt reagent kit with gdna eraser perfect real time, by Takara Bio (1 mentions) Anti collagen 4 ab6586, by Abcam (1 mentions) Anti flag sab4200071, by Merck Group (1 mentions) Nuclear and cytoplasmic protein extraction kit, by Keygen Biotech (1 mentions) Mtt assay kit, by Solarbio (1 mentions) Oil red o staining kit, by Solarbio (1 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) N formyl methionyl leucyl phenylalanine fmlf, by Merck Group (1 mentions) Fibronectin fn, by BD (1 mentions) Pluronic f 127, by Merck Group (1 mentions)

4 protocols using low endotoxin bovine serum albumin bsa

Murine Macrophage Fatty Acid Pretreatment

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Murine macrophage-like cell line RAW 264.7 cells (ATCC, Manassas, VA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Invitrogen, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS, endotoxin < 25 EU/mL; Sigma-Aldrich, St. Louis, MO), 100 U/mL of penicillin, and 100 μg/mL streptomycin (MP Biomedicals, LLC, Santa Anna, CA) at 37°C in a 5% CO₂ humidified incubator. Fatty acids were prepared from fatty acid sodium salts obtained from Nu-Check Prep, Inc., Elysian, MN (> 99% purity) with the exception of DHA which was obtained from Sigma-Aldrich (≥ 95% purity). Fatty acid sodium salts were combined with fatty acid-free, low endotoxin bovine serum albumin (BSA; Sigma-Aldrich) at a 2:1 molar ratio. Cells were pretreated with 100 μM of the fatty acid or BSA for 24 h. After fatty acid pretreatment, cells were stimulated with 100 ng/mL of ultra-pure LPS (Invivogen, San Diego, CA) of the E. coli 0111:B4 strain for the indicated times in DMEM containing 10% FBS in the presence or absence of the fatty acid/BSA complex. Cellular protein concentration was measured using the bicinchoninic acid method (Pierce Inc., Rockford, IL).

Honda K.L., Lamon-Fava S., Matthan N.R., Wu D, & Lichtenstein A.H. (2014). EPA and DHA exposure alters the inflammatory response but not the surface expression of toll-like receptor 4 in macrophages. Lipids, 50(2), 121-129.

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Sodium Palmitate-BSA Complex Preparation

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To make 0.1 M sodium palmitate (PA) solution, PA (0.0278 g; Sigma) was melted in 1 mL of 0.1 M NaOH solution at 70 °C. Low-endotoxin bovine serum albumin (BSA; 1.6 g; Sigma) was dissolved in 19 mL of DMEM or DMEM/F-12 medium with gentle shaking in a water bath at 37 °C for 1 h. The PA solution was added to the continuously vortexed BSA solution drop-by-drop. Then, the mixed solution was incubated in a water bath at 37 °C for another 30 min. The vehicle control was obtained by preparing an 8% BSA solution in DMEM or DMEM/F-12 medium. Both the BSA-PA solution and the vehicle control solution were filtered through a 0.22 μm syringe filter and stored at −20 °C until further study. The approximate BSA:PA molar ratio was 1:4.

Zhu W., Sahar N.E., Javaid H.M., Pak E.S., Liang G., Wang Y., Ha H, & Huh J.Y. (2021). Exercise-Induced Irisin Decreases Inflammation and Improves NAFLD by Competitive Binding with MD2. Cells, 10(12), 3306.

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Optimized Synthesis and Characterization of LM9

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LM9 was synthesized with a purity over 97% in our laboratory. PA was solubilized in low-endotoxin bovine serum albumin (BSA) (5%) at a final concentration of 5 mM. PA and BSA were purchased from Sigma-Aldrich (St. Louis, MO, USA). The MTT assay kit and oil red O staining kit (for cultured cells) were acquired from Solarbio (Beijing, China). Antibodies were purchased from the following suppliers: anti-lamin B1 (ab133741), anti-IκB alpha (ab133462), anti-Ly6G (ab25377), anti-TNF-α (ab6671), anti-collagen I (ab6308), and anti-collagen IV (ab6586) were obtained from Abcam (Cambridge, MA, USA). Anti-TLR4 (sc-293072) and anti-TGF-β (sc-130348) were obtained from Santa Cruz (Dallas, TX, USA). Anti-FLAG (SAB4200071) and anti-HA (MFCD00803873) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-ICAM-1 (Cat# 60299-1-Ig) was obtained from Proteintech (Wuhan, China). Two plasmids encoding pCMV-HA-MyD88 and Flag-MyD88 were obtained from Sino Biological Inc. (Beijing, China).
ELISA kits for mouse TNF-α, IL-1β, and IL-6 were purchased from Invitrogen (Carlsbad, CA, USA). Nuclear and cytoplasmic protein extraction kits were from KeyGEN Biotech (Nanjing, China). PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) was purchased from Takara Bio (RR047A, Shiga, Japan).

Zheng X.Y., Sun C.C., Liu Q., Lu X.Y., Fu L.L., Liang G., Zhang X.H, & Chen G.Z. (2020). Compound LM9, a novel MyD88 inhibitor, efficiently mitigates inflammatory responses and fibrosis in obesity-induced cardiomyopathy. Acta Pharmacologica Sinica, 41(8), 1093-1101.

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Fluorescent Protein Labeling Protocol

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Rinsing buffer was Hanks’ Balanced Salt Solution (Invitrogen, Carlsbad, CA) without calcium or magnesium supplemented with 10 mM HEPES (Invitrogen) and pH adjusted to 7.4. Storage buffer was rinsing buffer supplemented with 2 mg/mL glucose. Running buffer was storage buffer supplemented with 1.5 mM Ca²⁺ and 2 mM Mg²⁺. Fibronectin (FN) was from human plasma (BD Biosciences, Bedford, MA). Low-endotoxin bovine serum albumin (BSA) (Sigma) was prepared at 2% and 0.2% w/v in PBS without calcium and magnesium (PBS(−)). Labeling of proteins via Alexa Fluor carboxylic acid, succinimidyl ester (Invitrogen) was performed in accordance with the manufacturer’s recommended protocol. Stock N-formylmethionyl-leucyl-phenylalanine (fMLF) (Sigma, Saint Louis, MO) was reconstituted in glacial acetic acid before dilution. The nonionic triblock copolymer Pluronic F-127 (Sigma) was prepared at 0.2% w/v in PBS(−). All solutions were sterile filtered or prepared sterile. Bicinchoninic acid protein assays (Pierce Biotechnology, Rockford, Il) were performed on stock solutions of proteins to measure concentration.

Henry S.J., Crocker J.C, & Hammer D.A. (2014). Ligand density elicits a phenotypic switch in human neutrophils. Integrative biology : quantitative biosciences from nano to macro, 6(3), 348-356.

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