bicinchoninic acid
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Gels ◽  
2022 ◽  
Vol 8 (1) ◽  
pp. 43
Author(s):  
Dario Lucas Helbing ◽  
Leopold Böhm ◽  
Nova Oraha ◽  
Leonie Karoline Stabenow ◽  
Yan Cui

Despite the availability of a wide range of commercial kits, protein quantification is often unreliable, especially for tissue-derived samples, leading to uneven loading in subsequent experiments. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concentrations of tissue samples. We present a Ponceau S staining-based dot-blot assay as an alternative for protein quantification. This method is simple, rapid, more reliable than the BCA assay, compatible with biological samples lysed in RIPA or 2x SDS gel-loading buffer, and also inexpensive.


Bioanalysis ◽  
2021 ◽  
Author(s):  
Yong Jiang ◽  
Andrew P Mayer ◽  
Katie Carle ◽  
Reza Mozaffari ◽  
George Gunn

Aim: Ruthenium-labeled antibodies are commonly used detection reagents in bioanalysis assays and must be characterized to ensure quality. The aim of this work was to develop a method to determine the concentration and incorporation ratio (the degree of labeling [DOL]) of ruthenium-labeled antibodies by UV/VIS spectroscopy. Materials & methods: Free SULFO-TAG compound was scanned using UV/VIS and showed an absorbance peak at 292 nm. In contrast, antibodies demonstrate UV absorbance at 280 nm. After experimentally determining the extinction coefficients at 280 and 292 nm of free ruthenium and antibody, we generated a formula based on the Beer–Lambert law that calculates both concentration and DOL of these ruthenium-labeled antibodies. Conclusion: The concentration and DOL values determined by our method were comparable to those determined from bicinchoninic acid and LC/MS for the same reagents. This method creates a faster and more accessible reagent characterization process that uses far less reagent than the more traditional alternatives.


2020 ◽  
Vol 608 ◽  
pp. 113904 ◽  
Author(s):  
Javiera Cortés-Ríos ◽  
Ana María Zárate ◽  
Juan David Figueroa ◽  
Joaquín Medina ◽  
Eduardo Fuentes-Lemus ◽  
...  

2019 ◽  
Vol 43 (1) ◽  
pp. 277-283
Author(s):  
Yuma Saito ◽  
Michihiro Nishikawa ◽  
Taro Tsubomura

The repeatable colour change upon freezing–melting operation of a water solution of a Cu(i) complex is found. Efficient red light absorption is also one of the advantages of this complex.


Nano LIFE ◽  
2018 ◽  
Vol 08 (03) ◽  
pp. 1850005 ◽  
Author(s):  
Daniel L. Smith ◽  
Elizabeth N. Lemieux ◽  
Sutapa Barua

Conducting the bicinchoninic acid (BCA) assay directly after a coupling reaction using (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) (EDC) and [Formula: see text]-hydroxysuccinimide (NHS) chemistry produces significant errors. Here we present a correction for the quantification of gelatin in the supernatant (SN) following gelatin conjugation to polymer microparticles using EDC and NHS chemistry. Following the conjugation reaction, SNs from the gelatin-microparticle formation reaction are treated with BCA assay reagents and quantified for the percentage of unbound gelatin in the solution. NHS was found to interfere with the BCA assay reagents and is dependent on incubation time. It is found that the large concentration (500[Formula: see text][Formula: see text]g/mL) of NHS in the conjugation reaction interferes with the sensitivity of gelatin present in SNs. The interference from NHS requires a careful analysis to distinguish the BCA background absorbance from the sample absorbance. Using an NHS control solution can correct NHS interference and thus decrease the expensive iterations in gelatin quantification and enable accurate analysis of gelatin content. The accuracy of gelatin quantification is further improved by reducing the BCA assay incubation time to approximately 20[Formula: see text]min, compared with the recommended 30[Formula: see text]min. This re-assessment of BCA assay is important to avoid misestimating biases in bioconjugation processes.


BioTechniques ◽  
2018 ◽  
Vol 64 (6) ◽  
pp. 245-253 ◽  
Author(s):  
Beata A Butruk-Raszeja ◽  
Aleksandra Kuźmińska
Keyword(s):  

2018 ◽  
Vol 30 (3) ◽  
pp. 479-485
Author(s):  
Dinglong Chen ◽  
Xiaoying Wang ◽  
Yonghui He ◽  
Tianhan Kai ◽  
Zhiqiang Li ◽  
...  

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