Overview

  • Product name
    DyLight® 550 Conjugation Kit (Fast)
  • Product overview

    Dylight® 550 Fast Conjugation Kit (ab201800) uses a simple and quick process to conjugate an antibody to Dylight® 550. It can also be used to conjugate other proteins or peptides.


    To conjugate an antibody to Dylight® 550 using this kit:
    - add modifier to antibody and incubate for 15 mins
    - add quencher and incubate for 5 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 Dylight® 550.


     

  • Notes

    Amount and volume of antibody for conjugation to Dylight® 550

     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

    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 media are incompatible.

    DyLight® is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

     

  • Tested applications
    Suitable for: Conjugationmore details

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 30 µg 300 µg 1 mg
    Dylight® 550 Conjugation Mix 3 vials 3 vials 1 vial
    Dylight® 550 Modifier Reagent 1 vial 1 vial 1 vial
    Dylight® 550 Quencher Reagent 1 vial 1 vial 1 vial
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab201800 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 concentration.

Images

Protocols

References

ab201800 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
The conjugation kit was very easy to use and gave a very nice signal.
The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. Next, the cells were blocked in PBS / 10% normal goat serum / 0.3M glycine and subsequently incubated with Abcam Mouse monoclonal [3E5G11] to Villin - N-terminal (ab201989) conjugated with DyLight® 550 Fast Conjugation Kit (ab201800) (1/200) for 30 min at RT. Unlabelled sample (blue) was also used as a control.

Abcam user community

Verified customer

Submitted Sep 20 2018

Improved staining for mouse-on-mouse immunofluorescence

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Adult mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS.

Sections were then incubated with either:
(A) Mouse Anti-DDX4 (ab27591) at a 1:500 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Donkey Anti-Mouse 555 (ab150110) applied at a 1:500 dilution. (B) Mouse Anti-DDX4 (ab27591) labeled with Dylight 550 Fast Conjugation Kit (ab201800). Sections were incubated with labeled antibody at 1:100 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and mounted.

DDX4 is shown in red. Phalloidin, which stains F-actin is shown in blue.

Mr. Bryan Niedenberger

Verified customer

Submitted Sep 21 2016

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

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