bca reagent a and b composition

Figure 1. Albumin Standard (2 mg/ml): 10 × 1 ml ampules of bovine serum albumin (BSA) in 0.9% NaCl and 0.05% sodium azide. Store the Protein Standard at 2–8 °C. Pierce BCA Protein Assays have a unique advantage over the Coomassie dye–based assays (Bradford)–they are compatible with samples that contain up to 5% surfactants (detergents), and are affected much less by protein compositional differences, providing greater protein-to-protein uniformity. … >> This low protein-to-protein variation leads to higher accuracy in determining protein concentration for unknown protein samples. Pipette 50 µl of each protein … Proteins are diverse in their composition and structure, and with some assays the proteins’ differences in amino acid sequence, isoelectric point (pI), secondary structure, and side chains or prosthetic groups result in variation in the colorimetric response. 3. analytical standard (1) Electrophoresis (2) Molecular Biology Reagent (3) Reagent (1) Brand. Reagents and Equipment Required Depending on Assay Format Used • Spectrophotometer capable of measuring absorbance in the 595 nm region. Figure 4. 4. Figure 6. Both assays were conducted according to the respective manufacturers’ protocols, in a microplate format. For example, 5 M NaCl in a sample assayed by the standard assay is equivalent to 0.125 M NaCl in a sample assayed by the microassay … The reagent concentrations given in the reagent compatibility table are for the standard assay procedure (0.2–1.4 mg/ml protein). The standard Pierce BCA Protein Assay detects protein concentrations from 20 to 2,000 μg/mL using a two-component system: Reagent A, a carbonate buffer containing BCA reagent, and Reagent B, a cupric sulfate solution, which are combined to make an apple green–colored working solution that turns purple after 30 minutes at 37°C in the presence of protein. For the standard protocol, pipette the following into clean, dry test tubes: Standards/samples 100 ul, 500 ul reagent A (vortex), 4 ml reagent B (vortex). Create Account, A53225,A53226,23225,23227,23221,23228,23223,23222,23224,23230, Spectroscopy, Elemental & Isotope Analysis, Preclinical to Companion Diagnostic Development, Microbiological Media and Media Additives, Gel Electrophoresis Equipment and Supplies, BCA Protein Assay - Reducing Agent Compatible, User Guide: Pierce Rapid Gold BCA Protein Assay Kit, User Guide: Pierce BCA Protein Assay Kit − Reducing Agent Compatible, User Guide: Modified Lowry Protein Assay Kit, Application note: A rapid, sensitive protein assay for the accurate analysis of protein concentrations, Tech Tip: Determine acceptable wavelengths for measuring protein assays, Tech Tip: Eliminate interfering substances from samples for BCA Protein Assay, Tech Tip: How to use a protein assay standard curve, Tech Tip: Protein assay compatibility table, Less protein:protein variation than Coomassie dye methods, Significantly less (14–23%) protein:protein variation than Coomassie dye methods, Less protein:protein variation than BCA, Coomassie dye, or Lowry methods, Reducing reagents, Chelators, detergents, tris, tricine, glycerol. BCA Working Reagent (Reagent A mixed with Reagent B) is stable for one day. This … The variation as a function of protein composition could be decreased by reaction at 60°C. Although the mechanism of color formation for the Pierce Modified Lowry Protein Assay is similar to that of the Pierce BCA Protein Assay, there are several significant differences between the two. Add 500 microliters working solution to each 500 microliters sample … The sample is either a blank, a BSA … /Filter /FlateDecode The BCA Protein Assay is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantization of total protein. The copper complex exhibits a strong linear absorbance at 480 nm with increasing protein concentrations. The BCA– copper complex is water-soluble and exhibits a strong linear absorbance at 562 nm with increasing protein concentrations. 200 µl reagent B: Low-concentration assay: 5–250 µg/ml protein: 5–250 µg/ml protein : 200 µl sample: 20 µl sample : 100 µl reagent A: 10 µl reagent A : 800 µl reagent B: 80 µl reagent B: Compatible Reagents. 6) Prepare enough BCA assay reagents by combining 20 mL of Reagent A with 400 µL of Reagent B. The signal induced by the BCA reagent is approximately 100 times more sensitive (lower limit of detection) than the signal using the biuret reagent. If your lysis buffer contains detergent, add 20 ul of reagent S to each ml of reagent A. Concentrations compatible with the microassay procedure (1.25–25 µg/ml protein) are 1/40 of those listed. BCA-Arbeitslösung (BAL) Zur Herstellung der BAL mischen Sie 25 Teile Reagent A mit 25 Teilen Reagent B und 1 Teil Reagent C. Pro Ansatz werden 0,5 ml BAL benötigt, so dass die entsprechende Menge BAL zur Durchführung der Quantifizierung angesetzt werden sollte.

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