Overview

  • Product name
    FITC Conjugation Kit
    See all FITC kits
  • Product overview

    FITC Conjugation Kit ab102884 uses a simple and quick process to conjugate an antibody to FITC . It can also be used to conjugate other proteins or peptides.


    To conjugate an antibody to FITC using this kit:
    - add modifier to antibody and incubate for 3 hrs
    - add quencher and incubate for 30 mins
    The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.


    Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to FITC.

  • Notes

    Amount and volume of antibody for conjugation to FITC 

     Kit size   Recommended 
    amount of antibody
    1 
    Maximum 
    amount of antibody
    Maximum antibody 
    volume2
    30 µg   3 x 10 µg  3 x 20 µg 3 x 10 µL
    300 µg   3 x 100 µg  3 x 200 µg 3 x 100 µL
    1 mg   1 mg 2 mg 1 mL

    1 Using the maximum amount of antibody may result in less labelling per antibody.

    2 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 5mg/ml or < 0.5 mg/ml should be diluted /concentrated.

     

    Buffer Requirements for Conjugation

    Buffer should be pH 6.5-8.5.

    Compatible buffer constituents 
    If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.

    50mM / 0.6% Tris1 0.1%/1% BSA2 50% glycerol
    0.1% sodium azide PBS Potassium phosphate
    Sodium chloride HEPES Sucrose
    Sodium citrate EDTA Trehalose

    1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
    2 1% BSA gives lower quality conjugates, BSA can also interfere with the use of the conjugated antibody in tissue staining.


    Incompatible buffer constituents

    Thiomerosal Proclin Glycine
    Arginine Glutathione DTT

    If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.

    Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture 

  • Tested applications
    Suitable for: Conjugationmore details

Properties

Applications

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.

Application Abreviews Notes
Conjugation Use at an assay dependent dilution.

Images

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

Protocols

References

This product has been referenced in:
  • Li Q  et al. Megalin mediates plasma membrane to mitochondria cross-talk and regulates mitochondrial metabolism. Cell Mol Life Sci 75:4021-4040 (2018). Read more (PubMed: 29916093) »
  • Zhdanov DD  et al. Murine regulatory T cells induce death of effector T, B, and NK lymphocytes through a contact-independent mechanism involving telomerase suppression and telomere-associated senescence. Cell Immunol 331:146-160 (2018). Read more (PubMed: 29935763) »
See all 13 Publications for this product

Customer reviews and Q&As

Filter by Ratings

1-2 of 2 Abreviews

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.

Abcam user community

Verified customer

Submitted Nov 02 2015

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.

Hans Wilms

Verified customer

Submitted Jun 03 2014

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

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