Name: DNP-PEG4-NHS ester
Brand: BOC Sciences
Rating: 5 (1 reviews)

Synonyms

DNP-NH-PEG(4)-NHS; DNP-PEG4-NHS; 1-(2,4-Dinitrophenylamino)-3,6,9,12-tetraoxapentadecanoic acid succinimidyl ester; 2,5-dioxopyrrolidin-1-yl 1-((2,4-dinitrophenyl)amino)-3,6,9,12-tetraoxapentadecan-15-oate; 2,5-Pyrrolidinedione, 1-[3-[2-[2-[2-[2-[(2,4-dinitrophenyl)amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-oxopropoxy]-; 1-({15-[(2,4-Dinitrophenyl)amino]-4,7,10,13-tetraoxapentadecan-1-oyl}oxy)-2,5-pyrrolidinedione

IUPAC Name

(2,5-dioxopyrrolidin-1-yl) 3-[2-[2-[2-[2-(2,4-dinitroanilino)ethoxy]ethoxy]ethoxy]ethoxy]propanoate

Canonical SMILES

C1CC(=O)N(C1=O)OC(=O)CCOCCOCCOCCOCCNC2=C(C=C(C=C2)[N+](=O)[O-])[N+](=O)[O-]

InChI

InChI=1S/C21H28N4O12/c26-19-3-4-20(27)23(19)37-21(28)5-7-33-9-11-35-13-14-36-12-10-34-8-6-22-17-2-1-16(24(29)30)15-18(17)25(31)32/h1-2,15,22H,3-14H2

InChIKey

OZBOCCBKQAMCSA-UHFFFAOYSA-N

Density

1.4±0.1 g/cm3

Solubility

Soluble in DMSO

Appearance

Yellow Liquid

Shipping

Room temperature

Storage

Store at 2-8°C for short term (days to weeks) or -20°C for long term (months to years)

Boiling Point

686.3±65.0°C at 760 mmHg

DNP-PEG4-NHS ester, a versatile reagent in biochemistry and molecular biology, finds diverse applications. Explore four key applications of DNP-PEG4-NHS ester below:

Protein Labeling: Widely adopted for protein labeling, DNP-PEG4-NHS ester effectively attaches DNP (dinitrophenyl) groups to proteins for detection or purification purposes. This compound selectively reacts with amino groups on proteins, providing precise and simple labeling. Detection of DNP-labeled proteins can be achieved using anti-DNP antibodies, facilitating a range of biochemical assays with efficacy.

Antibody Production: In the realm of monoclonal and polyclonal antibody development, DNP-PEG4-NHS ester plays a pivotal role in creating antigen conjugates. Pairing DNP with a carrier protein stimulates robust immune responses, culminating in the generation of high-affinity anti-DNP antibodies. Such antibodies serve as invaluable assets for immunoassays and various research applications, elevating the arsenal of tools available to researchers.

Surface Functionalization: Delving into surface functionalization, DNP-PEG4-NHS ester emerges as a key player in enhancing microarrays and biosensors. By introducing DNP groups onto surfaces, researchers establish precise and reactive sites for subsequent biomolecule attachment. This sophisticated technique enables the development of highly specific and sensitive diagnostic platforms.

Studying Protein Interactions: Unlocking the realm of protein-protein and protein-ligand interactions, DNP-PEG4-NHS ester facilitates insightful studies. By tagging proteins with DNP, researchers can quantify interactions with other proteins or ligands using techniques like ELISA. This strategic approach aids in unraveling intricate molecular mechanisms and identifying potential drug targets, contributing to advancements in the field of protein interaction studies.

1.Binding mechanisms of PEGylated ligands reveal multiple effects of the PEG scaffold. Biochemistry

Das R, Baird E, Allen S, Baird B, Holowka D, Goldstein B

A series of synthetic ligands consisting of poly(ethylene glycol) (PEG), capped on one or both ends with the hapten 2,4-dinitrophenyl (DNP), were previously shown to be potent inhibitors of cellular activation in RBL mast cells stimulated by a multivalent antigen [Baird, E. J., Holowka, D., Coates, G. W., and Baird, B. (2003) Biochemistry 42, 12739-12748]. In this study, we systematically investigated the effect of increasing length of the PEG scaffold on the binding of these monovalent and bivalent ligands to anti-DNP IgE in solution. Our analysis reveals evidence for an energetically favorable interaction between two monovalent ligands bound to the same receptor, when the PEG molecular mass exceeds approximately 5 kDa. Additionally, for ligands with much higher molecular masses (>10 kDa PEG), the binding of a single ligand apparently leads to a steric exclusion of the second binding site by the bulky PEG scaffold. These results are further corroborated by data from an alternate fluorescence-based assay that we developed to quantify the capacity of these ligands to displace a small hapten bound to IgE. This new assay monitors the displacement of a small, receptor-bound hapten by a competitive monovalent ligand and thus quantifies the competitive inhibition offered by a monovalent ligand. We also show that, for bivalent ligands, inhibitory capacity is correlated with the capacity to form effective intramolecular cross-links with IgE.

2.Complement-mediated fragmentation of soluble and insoluble immune complexes containing porcine anti-DNP antibodies

Miklós K, Kulics J, Franĕk F, Füst G, Gergely J

Complement-mediated release of soluble immune complexes and immune precipitates containing DNP-PSA and precipitating or non-precipitating porcine anti-DNP antibodies was studied. A decrease in the average size of soluble immune complexes indicating their fragmentation was observed during incubation in excess human serum, the extent of the complex release was found to be in direct proportion to the time of incubation. The effect was complement-dependent. In the second part of the study, complement-dependent solubilization of the immune precipitates of the precipitating antibody preparation was compared to the solubilization of the precipitates of the non-precipitating antibody formed in the presence of PEG. Although, both types of precipitates activated complement in about the same extent, complexes of non-precipitating antibody were solubilized much slower than those of the precipitating one. As avidity of both antibody preparations to the antigen was high, the observed differences in the rate of the complex solubilization probably reflected differences in the structure of the two types of complexes.

3.Suppression of reaginic antibodies with modified allergens. IV. Induction of suppressor T cells by conjugates of polyethylene glycol (PEG) and monomethoxy PEG with ovalbumin

Lee WY, Sehon AH, Akerblom E

Administration of multiple injections of conjugates of ovalbumin (OA) and polyethylene glycol (PEG) or its monomethoxy derivative (mPEG) into mice which had been sensitized with 2,4-dinitrophenylated OA (DNP3-OA) abrogated both the anti-OA and anti-DNP IgE responses, in spite of additional injections of the sensitizing dose of DNP3-OA in the presence of A1(OH)3. Treatment of mice with OA-PEG in A1(OH)3 stimulated preferentially helper T cells, whereas injection of mice with OA-PEG in the absence of adjuvant elicited predominantly suppressor T cells. The unresponsive state of mice which had been treated 21 days earlier with OA-PEG could not be broken by the transfer of normal spleen cells and an additional sensitizing dose of DNP3-OA. Transfer of spleen cells from tolerized animals to normal mice dampened the capacity of the latter to mount both anti-DNP and anti-OA IgE responses; however, the suppressive effect of these cells was eliminated by treatment of the normal recipients with cyclophosphamide, which is a procedure known to inactive suppressor T cells, and hence it may be concluded that this effect was not due to the carryover of the tolerogen with the transferred cells. All these results provide strong support for the conclusion that the suppressor cells induced by the treatment of mice with OA-PEG and OA-mPEG conjugates belonged to a T cell subpopulation, and that the B cells of these mice were devoid of suppressive activity.