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FITC1

Sigma-Aldrich

FluoroTag FITC Conjugation Kit

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Synonym(s):
FITC
NACRES:
NA.32

packaging

pkg of (Kit contains reagents for 5 conjugations.)

Quality Level

fluorescence

λex 495 nm; λem 525 nm

storage temp.

2-8°C

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APOAFSAB3700662SAB3701046
vibrant-m

FITC1

FluoroTag FITC Conjugation Kit

fluorescence

λex 495 nm; λem 525 nm

fluorescence

-

fluorescence

-

fluorescence

-

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

Quality Level

200

Quality Level

300

Quality Level

100

Quality Level

100

Application

The Flourotag FITC Conjugation Kit can be used for Immunohistochemisty and Immunocytochemistry using Flow Cytometry. It is also be used for the conjugation of FITC to peptide hormones, cytokines, growth factors and proteins.

Features and Benefits

  • Suitable for both small (1 mg) and large (5 mg) scale conjugations
  • Completely aqueous procedure - no DMF needed
  • Fast gel filtration separation of conjugate from excess FITC
  • Complete protocols for conjugation and F/P ratio determination
  • Sufficient reagents for at least 5 conjugations of 5 mg protein each and for optimization of F/P ratio before scale-up
  • References for applications and protocols

Principle

Sigma offers a convenient kit for preparing FITC-labeled antibodies. Fluorescein isothiocyanate (FITC), Isomer 1, is a widely used fluorophore, popular because of its high quantum efficiency and stability when conjugated. FITC is yellow-orange in color with an absorption maximum at 495 nm. Upon excitation it emits a yellow-green color with an emission maximum at 525 nm. Conjugation occurs through free amino groups of proteins or peptides, forming a stable thiourea bond (see reaction). FITC conjugates of antibodies, lectins, hormones, and growth factors have been used in a variety of immunohistochemical and flow cytometry applications. The protocols have been optimized for antibodies, but may be adapted to other proteins by the end user.

Analysis Note

Procedure
1. Dissolve protein and FITC in carbonate-bicarbonate buffer.
2. Slowly add FITC to protein with stirring. Cover with foil and stir 2 hours at room temperature.
3. Separate conjugate from free FITC on G-25 column. Collect fractions.
4. Pool fractions containing conjugate.
5. Determine F/P ratio of conjugate spectrophotometrically.
6. Stabilize with 1% bovine serum albumin and 0.1% sodium azide and store at 0-5 °C.

Legal Information

FluoroTag is a trademark of Sigma-Aldrich Co. LLC

Kit Components Only

Product No.
Description

  • Sephadex G-25 column, 3.5 mL 1

  • Sephadex G-25 column, 9.1 mL 1

  • Fluorescein isothiocyanate isomer I - F7250KC-2MG 2 mg

Kit Components Also Available Separately

Product No.
Description
SDS

  • P3813Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions 5 pkgSDS

Related product

pictograms

Exclamation mark

signalword

Warning

hcodes

Hazard Classifications

Eye Irrit. 2

Storage Class

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable


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H Hidaka et al.
Journal of lipid research, 40(6), 1131-1139 (1999-06-05)
We previously reported the identity and purification of two HDL3-binding proteins in rat liver plasma membranes. As these proteins are candidate high density lipoprotein (HDL) receptors and probably multifunctional, including a role in HDL metabolism, we have considerable interest in
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Journal of controlled release : official journal of the Controlled Release Society, 238, 22-30 (2016-07-16)
Inertial cavitation mediated by ultrasound has been previously shown to enable skin permeabilisation for transdermal drug and vaccine delivery, by sequentially applying the ultrasound then the therapeutic in liquid form on the skin surface. Using a novel hydrogel dosage form
P Lloyd-Evans et al.
Transfusion medicine (Oxford, England), 9(2), 155-160 (1999-06-03)
The use of flow cytometry for quantifying fetomaternal haemorrhage is increasing, and has been shown to be more accurate than the Kleihauer-Betke test for evaluating larger bleeds of over 4 mL in volume. Red cells are stained with fluorescently labelled
P F Blackmore
Steroids, 64(1-2), 149-156 (1999-05-14)
Progesterone rapidly increased intracellular free calcium ([Ca2+]i) in human sperm, removal of extracellular Ca2+ prevented the increase in [Ca2+]i. The Ca2+ influx was not blocked by the T-type Ca2+ channel blocker mibefradil. However T-type calcium channels do appear to be

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