The gel was stained with SimplyBlue™ SafeStain protein stain using the microwave protocol to visualize the expressed proteins. 913, where C is concentration (mg/ml); A is absorbance at 280 nm; and D is dilution. CROSS-REFERENCE TO RELATED APPLICATIONS. Not for use in diagnostic procedures. Convenient - a ready-to-use formulation eliminates the need to heat, reduce, or add sample buffer prior to use. 5-fold among the proteins of the set. Examples of amino-reactive groups that can be present on a compound used to label lysine, histidine, tryptophan, or an N-terminal amino acid include, but are not limited to, isothiocyanates, isocyanates, acyl azides, N-hydroxysuccinimide (NHS) esters, haloacetyl compounds, maleimide derivatives, sulfonyl chlorides, aldehydes, ketones, glyoxals, epoxides, oxiranes, carbonates, aryl halides, imidoesters, carbodiimides, or acid anhydrides. 2-8) for reaction with thiol-reactive functional groups and carbonate or borate buffers (pH about 9) for reaction with isothiocyanates and dichlorotriazines. 10 ul Sharp Pre-stained Protein Standard formulation of Example 11 was run on a 4-12% acrylamide gradient Bis-Tris NuPAGE® gel run with 1×MES running buffer (Invitrogen, Carlsbad, Calif. After electrophoresis the gel was placed on a transparency having a copy of a measuring scale (FIG. The sample is vortexed for 10-15 seconds to disperse the pellet and then immediately mixed using a Polytron mixer. Novex sharp prestained protein ladder. The Novex Sharp Protein Standard is also available in an unstained format. 100 μl of 10 mg/ml lysozyme (Calbiochem, San Diego, Calif., USA) solution in water was brought up to a volume of 1 ml with a final concentration of 50 mM Tris pH=8 and 0.
Novex Sharp Prestained Protein Standard Curve
5 cm apart at the completion of electrophoresis. For example, the ratio of the number of residues of a target amino acid to molecular weight may be 4 residues per 10 kDa, or 0. In another example, an amino acid having a chemical group that behaves as a nucleophile at a pH lower than neutrality, for example, aspartate or glutamate, can be a target amino acid and one or more other amino acids that behaves as a nucleophile at a pH less than neutrality can be a non-target amino acid that is not present in a labeled protein standard or modified in a labeled protein standard. Synthesis of Red Dye #1 (8-Anilino-1-Naphthalenesulfonic Acid-Aminophenyl Vinyl Sulfone; 8-ANS-APVS). 1% SDS and then the sample was loaded. Novex sharp prestained protein standard.html. 5 kDa to greater than 250 kDa. In illustrative embodiments, the sequence lacks residues of a non-target amino acid.
Novex Sharp Prestained Protein Standard.Html
4 mM MgSO4; 220 μM dNTPs; and stabilizers; with the following primer sets: |50. Primer design allowed for each 50 kd TA clone to have unique sequence ends that facilitated vector construction as shown in Table 2. The amino acid sequence encoding the protein sequence can optionally be mutated to further reduce the number of residues of cysteine and/or other non-target amino acids, for example, histidine and/or tryptophan, which can be labeled in reactions that target lysine. A negative ion mode mass spectrum was obtained to be sure that a parent peak was seen at a mass to charge ratio of 492. The cell paste is vortexed for 10-20 seconds to break the pellet and the paste is mixed with the Polytron right away. Novex sharp prestained protein standard.com. Pre-labeled standards are labeled prior to separation or experimental procedures, and can be observed during or after separation procedures without performing additional steps required to stain the proteins in the midst of or at the conclusion of a separation or experimental procedure. For example, an engineered protein to be used for making pre-labeled protein standards can have one or more copies of an amino acid sequence with at least 70% or at least 80% identity with at least 20, at least 30, at least 40, or at least 50 contiguous amino acids of a thioredoxin sequence, in which lysine has been removed from the sequence by deletion or mutation of lysine codons in the nucleic acid sequence encoding the protein. The 1314 bp inserts (50 kDa) were gel purified on a 1. In preferred embodiments, the ratios of cysteine residues to molecule weight for the two or more, three or more, four or more, five or more cys-labeled proteins that lack lysine do not vary by more than 5%. The dye was eluted in acetonitrile and the colored fractions were collected. Data provided by: Qamar S, Cambridge Institute.
Invitrogen™ Novex™ Sharp Pre-stained Protein Standard. The width of bands visible to the naked eye from proteins having a molecular weight of at least 20 kDa to less than 100 kDa range in width from 0. 2-10HIS-PmeI clone B6 was digested with XhoI and PmeI. In some preferred embodiments, the selectively labeled proteins having a molecular weight of greater than 10 kDa or greater do not differ by more than 5% in their migration in denaturing acrylamide electrophoresis gels from the migration of the same proteins in unlabeled form. A protein standard selectively labeled on lysine is preferably labeled with a dye that comprises an sulfhydryl-reactive group. In certain embodiments, a labeling compound conjugated to a first amino acid is a dye. Malar J 19:367 (2020). Novex™ Sharp Pre-stained Protein Standard. A set of pre-labeled protein standards can comprise two or more labeled proteins, in which the two or more proteins comprise different numbers of copies of a sequence derived from a naturally-occurring protein, in which the number of residues of a non-target amino acid have been reduced relative to the naturally-occurring protein sequence. 8; Imidazole; 5M HCl; Cobalt II chloride. The calculated molecular weights of the proteins can be performed by curve-fitting of molecular weight to migration distances or point-to-point calculation. The label can be a chemiluminescent substance, where the output signal is generated by chemical modification of the signal compound; a metal-containing substance; or an enzyme, where there occurs an enzyme-dependent secondary generation of signal, such as the formation of a colored product from a colorless substrate.