概述

  • 产品名称FITC偶联试剂盒
    参阅全部 FITC 试剂盒
  • 产品概述

    Abcam's FITC Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with FITC. FITC is a green fluorescent dye which has an excitation wavelength of 495nm and an emission wavelength of 519nm.

    The conjugated antibody can be used straight away in WB, ELISA, IHC, Immunofluorescence (ICC/IF), Flow cytometry and FRET etc

    Learn more about buffer compatibility, protein/secondary antibody conjugation and labelling chemistry in our FAQs.

     

  • 说明

    The antibody to be labelled should be purified, in an appropriate buffer for conjugation and at a suitable concentration, as described in the protocol booklet. If not, consider using our antibody purification and concentration kits.

     

    Pre-conjugation considerations

    The purified antibody to be labeled should ideally be in 10-50mM amine-free buffer (e.g. MES, MOPS, HEPES, PBS), pH range 6.5 to 8.5. If the buffer is more concentrated or outside this pH range contact the technical support team.

    Common non-buffering salts (e.g. sodium chloride), chelating agents (e.g. EDTA), and sugars have no effect on conjugation efficiency. Azide (0.02 to 0.1%) and BSA (0.1 to 0.5%) have little or no effect. Glycerol up to 50% has no effect.

    Avoid buffer components that are nucleophilic, as these may react with the kit chemicals. Primary amines (e.g. amino acids or ethanolamine) and thiols (e.g. mercaptoethanol or DTT) fall within this class (Note: Tris-based buffers should be avoided). If your buffer contains primary amines and/or thiols, you should consider using our Concentration and Purification Kits (ab102778 or ab102784).

    Recommended amount and volume of antibody for optimal results

    Pack sizeAmount of antibodyVolume of antibody
    Up to 20 µg10-20 µg4-10 µl
    Up to 200 µg100-200 µg40-100 µl
    Up to 2 mg1-2 mg400-1000 µl

    Antibody concentrations of 1-4 mg/ml generally give optimal results. For conditions outside the suggested ranges contact the technical support team.

    Conjugation reaction

    • Before adding the antibody to the FITC mix, add 1 µl of Modifier reagent to each 10 µl of antibody to be labelled. Mix gently.
    • Remove cap from vial of FITC mix and pipette the antibody sample (with added  Modifier) directly onto the lyophilized material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.

     

    • Replace cap on the vial and leave standing in the dark at room temperature (20-25°C) for 3 hours. Conjugations can also be set up and left overnight; longer incubation times have no negative effect on the conjugate.

     

    • After incubating for 3 hours (or more), add 1 µl of  Quencher  reagent for every 10 µl of antibody used and mix gently. The conjugate can be used after 30 minutes. The conjugates do not require purification.

     

    Storage of conjugates

    Storage at 4°C is recommended for any conjugate. A preservative may be desirable for long-term storage. Other storage conditions may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.

    Disclaimer

    Labeling of the antibody will not work if the conjugation blocks the active paratope. This situation is rare.

  • 经测试应用适用于: Conjugationmore details

性能

  • 别名
    • Fluorescein
    • Fluorescein isothiocyanate

应用

Our Abpromise guarantee covers the use of ab102884 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Conjugation Use at an assay dependent dilution.

FITC Conjugation Kit 图像

  • This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.

实验方案

FITC Conjugation Kit (ab102884)参考文献

This product has been referenced in:
  • MacKrell JG  et al. Molecular targets of androgen signaling that characterize skeletal muscle recovery and regeneration. Nucl Recept Signal 13:e005 (2015). Read more (PubMed: 26457071) »
  • Isgrò M  et al. The C-C motif chemokine ligands CCL5, CCL11, and CCL24 induce the migration of circulating fibrocytes from patients with severe asthma. Mucosal Immunol 6:718-27 (2013). Read more (PubMed: 23149666) »

See all 6 Publications for this product

Product Wall

FITC Conjugated full length EpCAM Protein

Excellent Excellent 5/5 (Ease of Use)
Abreviews
FITC was succesfully Conjugated with Human EpCAM Protein. It was quite useful.
Username

Abcam user community

Verified customer

提交于 Nov 02 2015

There is no difference in sensitivity between the rapid (eg. ab188285) and the standard (eg. ab102884) labeling kits. The only difference between the two is the incubation time required for the reaction.

Better results than with secondary antibody.

Excellent Average 3/5 (Ease of Use)
Abreviews
The kit was easy enough for me to hand this off to an undergraduate, albeit with some explanation of the directions. These were a tad bit confusing at first, and not really clear on the volume of antibody to add (they give you a couple options since the Ab concentration of course matters, but didn't make it clear they were different options).
Otherwise the kit work fantastically with some store bought, pure, concentrated antibody! It actually gave better results than our previous method of using a conjugated secondary antibody when used in Flow Cytometry after doing an Intracellular Stain.
Username

Hans Wilms

Verified customer

提交于 Jun 03 2014


This antibody will detect the whole cells, and because the antiserum has not been absorbed (only Protein A purified) it may react with related microorganisms.


As this antibody is suitable for conjugation purpose according to the data...

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Thank you for your enquiry.

The EasyLink conjugation kits are designed so that they can be used to label proteins other than antibodies. In this case you should be able to label the Fc fused protein in the same way as labeling an antibody. H...

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Thank you for contacting us.

The labeling kit works by targeting free amines groups on your antibody. We therefore cannot guarantee that the label will not bind to the FAB area, however this would be the case for any other amine orientated lab...

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Thank you for your inquiry and your interest in our products.

I will have to double-check with the lab regarding glycosylation of the antibodies since I do not have this information readily at hand. I will let you know once I have received th...

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Thank you for your interest in the FITC conjugation kit. Unfortunately, it will be difficult to achieve a consistent ratio of FITC : IgG without extensive testing of the kit to determine the appropriate conditions. We have not done this testing, as the...

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Thank you for contacting us.

The fluorescein in ourKit is not simply fluorescein and it is not FITC either. Our fluorescein has been specially treated with proprietary Easy-Link chemistry to make the unique one-step conjugation possible.
<...

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Thank you for your inquiries.

I can confirm that additives in the buffer of the antibody will interfere with the labeling reaction.

We offer a kit that will concentrate the antibodyand also remove the additives:

ab102776 <...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"