Name: Fluorescein NHS Antibody Labeling Kit (1 x 1 mg)
Brand: BroadPharm
SKU: BP-50012
Price: 380.00 USD
Availability: InStock

Fluorescein NHS Antibody Labeling Kit

Catalog #: bp-50012

HeLa cells fixed with 4% paraformaldehyde and permeabilized with .1%. Specimen was incubated with Mouse Anti β-Tubulin IgG.

Key Features and Details

The HeLa cells were fixed with 4% paraformaldehyde, and permeabilized with 0.1% Triton X-100. Then, the specimen was blocked in BSA/PBS blocking buffer. Subsequently, the specimen was incubated with Mouse Anti β-Tubulin IgG, and, after washing, was further incubated with biotin-conjugated Goat Anti-Mouse IgG Biotin Conjugate. Finally, it was rinsed with PBS and stained with Streptavidin FITC Conjugate at 5 μg/mL (shown in green) and DAPI-2HCl at 1 μg/mL (shown in blue)

Overview
The fluorescein labeling kits are high-performance derivatives of fluorescein dye, activated for easy and reliable labeling of antibodies, proteins and other molecules for use as fluorescent probes.
Fluorescein dyes are used in wide-ranging applications including fluorescence microscopy, flow cytometry and immunofluorescence-based assays such as Western Blotting and ELISA. Fluorescein NHS are reactive toward primary amine groups on proteins, peptides and other biomolecules, while fluorescein--maleimide react with free sulfhydryl on cysteine residues.

Most activated fluorescein derivatives are mixtures of isomers with reactive groups attached at the 5- and 6-positions of the bottom ring. There is a big difference in flow cytometry although it is a small difference in the properties of these isomers in terms of excitation and emission spectra. The small difference will make your assay complicated because the ratio of two isomers varies lot by lot.
BroadPharm fluorescein labeling kits use single isomer- 6-isomer NHS and Maleimade to solve the issues.
- Amine and thio-specific labeling: the NHS-ester and maleimide efficiently labels antibodies and other purified proteins at primary amines or thiol.
- Optimized procedure: following the standard protocol results in antibodies with excellent dye:protein ratios for optimum activity and fluorescence
- Single-use fluors: no need to weigh tiny amounts powder; kits contain single-use vials of reagent
- Efficient purification: kits include purification resin and easy-to-use spin columns, ensuring rapid and efficient removal of non-reacted dye and excellent protein recovery.
* single isomer, no lot-to-lot variability.
Components
1. Dye NHS
2. Reaction buffer
3. Desalt column
4. DMSO, 1 rxn 1 mL
5. NaN3 3%,
Application
Fluorescein is mainly used to label primary antibody and secondary antibody. The application
- FC (981)
- ICFC (139)
- ICC (6)
- IHC-F (9)
- IHC-P (6)
- IF (3)
- IP (2)
- WB (2)
Images

The HeLa cells were fixed with 4% paraformaldehyde, and permeabilized with 0.1% Triton X-100. Then, the specimen was blocked in BSA/PBS blocking buffer. Subsequently, the specimen was incubated with Mouse Anti β-Tubulin IgG, and, after washing, was further incubated with biotin-conjugated Goat Anti-Mouse IgG Biotin Conjugate. Finally, it was rinsed with PBS and stained with Streptavidin FITC Conjugate at 5 μg/mL (shown in green) and DAPI-2HCl at 1 μg/mL (shown in blue)

CWR22R cells were fixed with 4% paraformaldehyde, blocked by 3% BSA. Subsequently the specimen was cultured at 4°C overnight with Anti PSMA monoclonal antibodies (1 : 100), and incubated with FITC-conjugated anti mouse IgG antibody at room temperature for 2h. DAPI nucleic stain was performed in blue.

Flow cytometry surface staining pattern of human peripheral whole blood stained using anti-human CD193 (5E8) FITC antibody prepared by MagicLink™ Fluoresein NHS kit ( Cat. # BP-50012)(4 µl reagent / 100µl of peripheral whole blood).
Protocol

FITC General protocol dye MAL labeling kit General protocol dye NHS labeling kit
Reference
1. Dolina JS, Lee J, Griswold RQ, et al.; TLR9 Sensing of Self-DNA Controls Cell-Mediated Immunity to Listeria Infection via Rapid Conversion of Conventional CD4+ T Cells to Treg.; Cell Rep.; 1(31) 2020 107249
2. Daniel et al.; miR-9a mediates the role of Lethal giant larvae as an epithelial growth inhibitor in Drosophila.; 2018;Biology;
3. Bankoti et al. Differential regulation of Effector and Regulatory T cell function by Blimp1.; 2017; Scientific Reports;
4. Herr AE et al; On-chip coupling of isoelectric focusing and free solution electrophoresis for multidimensional separations, Anal Chem (2003) 75:1180-1187
5. Knight SC et al, Transfer of antigen between dendritic cells in the stimulation of primary T cell proliferation; Eur J Immunol (1998) 28:1636-1644
6. The TH, Feltkamp TE. Conjugation of fluorescein isothiocyanate to antibodies. II. A reproducible method. Immunology. 1970;18(6):875-881.

Fluorescein NHS Antibody Labeling Kit

Fluorescein NHS Antibody Labeling Kit

Key Features and Details

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Overview

Components

Application

Images

Protocol

Reference

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