Intestinal tissue was prepared for ELISA as described previously [27 (link)]. Intestinal tissue lysates and luminal contents were evaluated by ELISA for CXCL1 (R&D Systems), TNF-α (eBioscience), myeloperoxidase (MPO; R&D Systems) [28 (link)], and albumin (Bethyl Laboratories) according to the manufacturers’ instructions. Total protein concentration was measured using the Pierce BCA Protein Assays Kit (Thermo Scientific).
Pierce bca protein assays kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Pierce BCA Protein Assays Kit is a colorimetric detection kit used to quantify the total protein concentration in a sample. The kit utilizes the bicinchoninic acid (BCA) reaction, where proteins reduce Cu2+ to Cu+ in an alkaline environment, and the resulting Cu+ ions chelate with BCA to produce a purple-colored complex that absorbs light at 562 nm. The intensity of the color is proportional to the protein concentration, allowing for accurate quantification.
Automatically generated - may contain errors
Lab products found in correlation
Myeloperoxidase mpo, by R&D Systems (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Cxcl1, by R&D Systems (1 mentions)
Protease inhibitor and phosphatase inhibitor cocktail, by Roche (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence substrate, by PerkinElmer (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
0.45 μm filter, by Merck Group (1 mentions)
3 protocols using pierce bca protein assays kit
1
Intestinal Tissue Preparation for ELISA Analysis
2
Tumor Protein Extraction and Western Blot
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Fresh or frozen tumor samples were mechanically chopped into small pieces and erythrocytes were removed. The preparation was then sonicated in cell lysis buffer (Cell Signaling Technology) with an EDTA-free protease inhibitor cocktail and phosphatase inhibitors (Roche). After brief sonication, the tumor lysates were centrifuged, and the supernatants were used as cell lysates. After that, the protein concentration was measured using a Pierce™ BCA protein assays kit (Thermo Fisher Scientific). Proteins were resolved on 10% precast gels (Invitrogen) and transferred to a nitrocellulose membrane. After blocking with 5% non-fat milk in Tris-buffered saline with 0.2% Triton X-100, membranes were incubated with the cleaved caspase 3 (Cell Signaling Technology) or GAPDH (Cell Signaling Technology) antibody, followed by the secondary antibody conjugated with horseradish peroxidase. After washing, the bands were visualized with enhanced chemiluminescence substrate (Perkin Elmer) and quantified using Image Quant LAS4000 (GE Healthcare).
3
Mucus Collection and Purification from Manila Clams
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Mucus was aspirated from the body surface (mantle and branchia) of non-anaesthetised Manila clam adults (n = 61) using a disposable Pasteur pipette. Mucus was sonicated by two cycles of 30 s at low intensity, centrifuged at 10,000× g for 15 min at 4 °C and filtered through a 0.45 μM filter (Millipore, Burlington, MA, USA) to remove debris. Protein concentration was measured using a Pierce BCA Protein assays kit (Thermo Fisher Scientific, Waltham, MA, USA) and mucus was collected and stored at −80 °C until use.
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