Product uses include measuring protein in
The Precision Red Advanced Protein Assay Reagent is designed to optimize the speed and accuracy of protein measurement. The reagent combines the useful properties of low protein to protein variance (See Fig 1) an d a wide protein concentration for a robust assay. The assay can measure accurately protein concentrations ranging from 0.25 to 50 mg/ml and is detergent compatible. A simple one step procedure results in a red to blue color change within 1 min, which can be recognized by measuring absorbance at 600 nm.
This reagent is so reproducible with a wide range of proteins and protein concentrations that it is employed to accurately determine the concentration in all of Cytoskeleton, Inc's protein products.
The Precision Red Advanced Protein Assay makes it very easy and quick to measure prottein concentrations. A general method looks as follows:
Detergent and chemical compatibility
The Precision Red Advanced Protein Assay is a highly stable assay and it tolerates a wide range of reagents. See the Advanced Protein Assay reagent compatibility page for a full list of reagents that have been tested.
Using the Precision Red Advanced Protein Assay in cell extracts
Since the Precision Red Advanced Protein Assay is highly compatible with non-ionic detergents (see the ADV Chemical tolerance page), this reagent is ideal for measuring protein concentrations in cell and tissue extracts. Furthermore, since protein concentration can be measured after only 1 min of incubation with the reagent, it allows the user to proceed with the experiment quicker than any other protein assay, something that can be of big importance when working with labile cell extracts.
Cytoskeleton, Inc. recommends using ADV02 for all our kits and assays that involve cell or tissue lysis. In the G-LISA™ small G-protein activation kits, we have even included this reagent to ensure that the user gets the most accurate and quickest possible measurement of the protein concentration in the cell lysates generated.
|Cruz-Nova, Pedro et al.||131I-C19 Iodide Radioisotope and Synthetic I-C19 Compounds as K-Ras4B–PDE6δ Inhibitors: A Novel Approach against Colorectal Cancer—Biological Characterization, Biokinetics and Dosimetry||Molecules 2022, Vol. 27, Page 5446||2022||ISSN 1420--3049|
|Xu, Weiyi et al.||Paxillin promotes breast tumor collective cell invasion through maintenance of adherens junction integrity||Molecular Biology of the Cell||2022||ISSN 1939-4586|
|Shields, Mario A. et al.||Gα13 loss in Kras/Tp53 mouse model of pancreatic tumorigenesis promotes tumors susceptible to rapamycin||Cell Reports||2022||ISSN 2211-1247|
|Hassin, Ori et al.||Different hotspot p53 mutants exert distinct phenotypes and predict outcome of colorectal cancer patients||Nature Communications 2022 13:1||2022||ISSN 2041--1723|
|Xu, Weiyi et al.||Paxillin promotes breast tumor collective cell invasion through maintenance of adherens junction integrity||Molecular biology of the cell||2022||ISSN 1939-4586|
|Datta, Gaurav et al.||Endolysosome Localization of ERα Is Involved in the Protective Effect of 17α-Estradiol against HIV-1 gp120-Induced Neuronal Injury||The Journal of neuroscience : the official journal of the Society for Neuroscience||2021||ISSN 1529-2401|
|Rojas-Colón, Luis A. et al.||4R-cembranoid confers neuroprotection against LPS-induced hippocampal inflammation in mice||Journal of Neuroinflammation||2021||ISSN 1742-2094|
|Dolmatova, Elena V et al.||Endothelial Poldip2 regulates sepsis-induced lung injury via Rho pathway activation||Cardiovascular Research||2021||ISSN 0008--6363|
|Liao, Liang et al.||Disrupting RhoA activity by blocking Arhgef3 expression mitigates microglia-induced neuroinflammation post spinal cord contusion||Journal of Neuroimmunology||2021||ISSN 1872-8421|
|Nehme, Jamil et al.||High dietary protein and fat contents exacerbate hepatic senescence and SASP in mice||FEBS Journal||2021||ISSN 1742-4658|
|Le, Anh Hoang et al.||Cyri-a limits invasive migration through macropinosome formation and integrin uptake regulation||Journal of Cell Biology||2021||ISSN 1540-8140|
|Hryniewicz-Jankowska, Anita et al.||Article the effect of neddylation inhibition on inflammation-induced mmp9 gene expression in esophageal squamous cell carcinoma||International Journal of Molecular Sciences||2021||ISSN 1422-0067|
|Papalazarou, Vassilis et al.||The creatine–phosphagen system is mechanoresponsive in pancreatic adenocarcinoma and fuels invasion and metastasis||Nature Metabolism||2020||ISSN 2522-5812|
|Bordeleau, Francois et al.||Tissue transglutaminase 2 regulates tumor cell tensional homeostasis by increasing contractility||Journal of Cell Science||2020||ISSN 1477-9137|
|Dias Gomes, Martim et al.||Polarity signaling ensures epidermal homeostasis by coupling cellular mechanics and genomic integrity||Nature Communications||2019||ISSN 2041-1723|
|Cederquist, Gustav Y. et al.||Specification of positional identity in forebrain organoids||Nature Biotechnology||2019||ISSN 1546-1696|
|Umesalma, Shaikamjad et al.||RABL6A inhibits tumor-suppressive PP2A/AKT signaling to drive pancreatic neuroendocrine tumor growth||Journal of Clinical Investigation||2019||ISSN 1558-8238|
|Paonessa, Francesco et al.||Microtubules Deform the Nuclear Membrane and Disrupt Nucleocytoplasmic Transport in Tau-Mediated Frontotemporal Dementia||Cell Reports||2019||ISSN 2211-1247|
|Kikuchi, Daniel S. et al.||Poldip2 mediates blood-brain barrier disruption in a model of sepsis-associated encephalopathy||Journal of Neuroinflammation||2019||ISSN 1742-2094|
|Tedeschi, Andrea et al.||ADF/Cofilin-Mediated Actin Turnover Promotes Axon Regeneration in the Adult CNS||Neuron||2019||ISSN 1097-4199|
|Stephens, Matthew et al.||Mesenteric lymphatic alterations observed during DSS induced intestinal inflammation are driven in a TLR4-PAMP/DAMP discriminative manner||Frontiers in Immunology||2019||ISSN 1664-3224|
|Juin, Amelie et al.||N-WASP Control of LPAR1 Trafficking Establishes Response to Self-Generated LPA Gradients to Promote Pancreatic Cancer Cell Metastasis||Developmental Cell||2019||ISSN 1878-1551|
|Zhang, Xiaoqing et al.||Alterations of MEK1/2-ERK1/2, IFNγ and Smad2/3 associated Signalling pathways during cryopreservation of ASCs affect their differentiation towards VSMC-like cells||Stem Cell Research||2018||ISSN 1876-7753|
|Borin, Mirta et al.||Rac1 activation links tau hyperphosphorylation and Aß dysmetabolism in Alzheimer's disease||Acta Neuropathologica Communications||2018||ISSN 2051-5960|
|Casique-Aguirre, Diana et al.||KRas4B-PDE6δ complex stabilization by small molecules obtained by virtual screening affects Ras signaling in pancreatic cancer 06 Biological Sciences 0601 Biochemistry and Cell Biology||BMC Cancer||2018||ISSN 1471-2407|
|Zhu, Yili et al.||An in vitro Microscopy-based Assay for Microtubule-binding and Microtubule-crosslinking by Budding Yeast Microtubule-associated Protein||Bio-Protocol||2018||ISSN 2331--8325|
|Proto, Jonathan D. et al.||Regulatory T Cells Promote Macrophage Efferocytosis during Inflammation Resolution||Immunity||2018||ISSN 1097-4180|
|Mangiatordi, Giuseppe Felice et al.||Automated identification of structurally heterogeneous and patentable antiproliferative hits as potential tubulin inhibitors||Chemical Biology and Drug Design||2018||ISSN 1747-0285|
|Meiring, Joyce C.M. et al.||Co-polymers of Actin and Tropomyosin Account for a Major Fraction of the Human Actin Cytoskeleton||Current Biology||2018||ISSN 0960-9822|
|Cruz-Nova, Pedro et al.||The small organic molecule C19 binds and strengthens the KRAS4b-PDEδ complex and inhibits growth of colorectal cancer cells in vitro and in vivo 11 Medical and Health Sciences 1112 Oncology and Carcinogenesis||BMC Cancer||2018||ISSN 1471-2407|
|Alhasani, Reem Hasaballah et al.||Gypenosides protect retinal pigment epithelium cells from oxidative stress||Food and Chemical Toxicology||2018||ISSN 1873-6351|
|McQueeney, Kelley E. et al.||Targeting ovarian cancer and endothelium with an allosteric PTP4A3 phosphatase inhibitor||Oncotarget||2018||ISSN 1949-2553|
|Patnaik, Sarita Rani et al.||RPGR protein complex regulates proteasome activity and mediates store-operated calcium entry||Oncotarget||2018||ISSN 1949-2553|
|Horita, Henrick et al.||Utilizing a comprehensive immunoprecipitation enrichment system to identify an endogenous post-translational modification profile for target proteins||Journal of Visualized Experiments||2018||ISSN 1940-087X|
|Mangiatordi, Giuseppe Felice et al.||Novel chemotypes targeting tubulin at the colchicine binding site and unbiasing P-glycoprotein||European Journal of Medicinal Chemistry||2017||ISSN 1768-3254|
|Huang, Jia Lin et al.||Lipoprotein-biomimetic nanostructure enables efficient targeting delivery of siRNA to Ras-activated glioblastoma cells via macropinocytosis||Nature Communications||2017||ISSN 2041-1723|
|Albalawi, Aishah et al.||Protective effect of carnosic acid against acrylamide-induced toxicity in RPE cells||Food and Chemical Toxicology||2017||ISSN 1873-6351|
|Hendricks, Louie et al.||Milk fat globule-epidermal growth factor-factor VIII–derived peptide MSP68 is a cytoskeletal immunomodulator of neutrophils that inhibits Rac1||Journal of Surgical Research||2017||ISSN 1095-8673|
|Chen, Weihua et al.||High RhoA expression at the tumor front in clinically localized prostate cancer and association with poor tumor differentiation||Oncology Letters||2016||ISSN 1792-1082|
|Lampi, Marsha C. et al.||Simvastatin ameliorates matrix stiffness-mediated endothelial monolayer disruption||PLoS ONE||2016||ISSN 1932-6203|
|Chen, Lihua et al.||Smooth muscle-alpha actin inhibits vascular smooth muscle cell proliferation and migration by inhibiting rac1 activity||PLoS ONE||2016||ISSN 1932-6203|
|Zan, G. Y. et al.||p38 mitogen-activated protein kinase activation in amygdala mediates κ opioid receptor agonist U50,488H-induced conditioned place aversion||Neuroscience||2016||ISSN 1873-7544|
|Gil-Henn, H. et al.||Arg/Abl2 promotes invasion and attenuates proliferation of breast cancer in vivo||Oncogene||2013||ISSN 0950-9232|
|Zilberter, Misha et al.||Dietary energy substrates reverse early neuronal hyperactivity in a mouse model of Alzheimer's disease||Journal of Neurochemistry||2013||ISSN 1471--4159|
|Takefuji, Mikito et al.||RhoGEF12 controls cardiac remodeling by integrating G protein- and integrindependent signaling cascades||Journal of Experimental Medicine||2013||ISSN 0022-1007|
|Castillo-Pichardo, Linette et al.||Dietary grape polyphenol resveratrol increases mammary tumor growth and metastasis in immunocompromised mice||2013||PMID 23298290|
|Padró, Teresa et al.||LDL-induced impairment of human vascular smooth muscle cells repair function is reversed by HMG-CoA reductase inhibition||PLoS ONE||2012||ISSN 1932-6203|
|Liu, Yao et al.||Actin polymerization-dependent increase in synaptic Arc/Arg3.1 expression in the Amygdala is crucial for the expression of aversive memory associated with drug withdrawal||Journal of Neuroscience||2012||ISSN 0270-6474|
|Ray, Anasuya et al.||Hedgehog signaling pathway regulates the growth of ovarian cancer spheroid forming cells||International Journal of Oncology||2011||ISSN 1019-6439|
|Hook, Jeff et al.||Functional identity of the gamma tropomyosin gene: Implications for embryonic development, reproduction and cell viability||BioArchitecture||2011||ISSN 1949-100X|
|Aguilar, Hector N. et al.||Quantification of rapid myosin regulatory light chain phosphorylation using high-throughput in-cell western assays: Comparison to western immunoblots||PLoS ONE||2010||ISSN 1932-6203|
|Romero, Ana M. et al.||Chronic ethanol exposure alters the levels, assembly, and cellular organization of the actin cytoskeleton and microtubules in hippocampal neurons in primary culture||Toxicological Sciences||2010||ISSN 1096-6080|
Question 1: Can I change the dilution volume of the experimental lysate being assayed?
Answer 1: Yes, different volumes of lysate can be used to quantify protein levels. The equation to determine protein concentration in µg/ml is: OD reading x dilution factor x 100 µg/ml/cm if a small volume (1.0 ml) cuvette is used. If a 96 well plate is used, the same equation is used with this single modification: 125 µg/ml/cm is the value used rather than 100 µg/ml/cm.
Question 2: Can this assay be performed using 96 well plates?
Answer 2: Yes, the protein assay can be performed in either small volume cuvettes (1.0 ml) or a 96 well plate. If a 96 well plate is used, the equation to determine protein concentration in µg/ml is: OD reading x dilution factor x 125 µg/ml/cm.
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