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    products/conjugation-kits/dylight633-conjugation-kit-fast-lightning-link-ab201802.pdf

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DyLight® 633 Conjugation Kit (Fast) - Lightning-Link® (ab201802)

  • Datasheet
  • SDS
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Immunohistochemistry (Frozen sections) - DyLight633 Conjugation Kit (Fast)- Lightning-Link (ab201802)
  • conjugation

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Overview

  • Product name

    DyLight® 633 Conjugation Kit (Fast) - Lightning-Link®
  • Product overview

    DyLight® 633 Conjugation Kit / DyLight® 633 Labeling Kit ab201802 uses a simple and quick process  for DyLight 633 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.


    To conjugate an antibody to DyLight® 633 using this kit:
    - add modifier to antibody and incubate for 15 mins
    - add quencher and incubate for 5 mins
    The DyLight® 633 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® 633.


    Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.

  • Notes

    This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid DyLight® 633 Labeling Kit. 325-0015 is the same as the 1 mg size. 325-0010 is the same as the 3 x 100 ug size. 325-0030 is the same as the 3 x 10 ug size. 325-0005 is the same as the 100 ug size.

    Amount and volume of antibody for conjugation to DyLight® 633 

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

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

    2 Ideal antibody concentration is 1 mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2 mg/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% 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 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.

    Storing and handling conjugation kits

    Lyophilized Lightning-Link® components are hygroscopic.

    Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.

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

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg
    ab274022 - Dylight&reg; 633 Conjugation Mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg
    ab273994 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
    ab273995 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Conjugation Kits
    • Dylight

Associated products

  • Related Products

    • Antibody Concentration And Clean-Up Kit (ab102778)
    • Antibody Purification Kit (Protein A) (ab102784)
    • Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)

Images

  • Immunohistochemistry (Frozen sections) - DyLight633 Conjugation Kit (Fast)- Lightning-Link (ab201802)
    Immunohistochemistry (Frozen sections) - DyLight633 Conjugation Kit (Fast)- Lightning-Link (ab201802)Image from Liu, Mengrui, et al., Adv Sci., 8(1):2002127; doi: 10.1002/advs.202002127. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Liu, Mengrui, et al used DyLight® 633 Conjugation Kit (Fast) - Lightning-Link® (ab201802) as part of examining Biodistribution of inhaled PT?BsAbs. They used the kit to conjugate DyLight® 633 to CD34-CD42b platelet?targeting bispecific antibodies (PT?BsAbs) for use in immunohistochemistry (Frozen sections).
    a-d) Representative confocal images showing distribution of PT?BsAbs in the bronchi and parenchyma of MI mice, respectively, a,b) 1 h and c,d) 6 h after inhalation. PT?BsAbs were pre-labeled with DyLight 633 (gray). Lung cells were stained with AF 594?labeled Phalloidin antibody (green), lung vasculature was stained by von Willebrand Factor (vWF) antibody (red); nuclei were stained with DAPI (blue). Scale bar, 100 µm. e) Quantification of PT BsAbs in MI mice in the lung 1 h and 6 h after inhalation, respectively. f) Time?course quantification results of PTBsAbs in MI mice in vasculature of lung 1 h and 6 h after inhalation, respectively. Confocal imaging revealing the PT BsAbs retention in the border zone of MI heart 6 h after g) inhalation or h) i.v. injection. Cardiomyocytes were stained with alpha sarcomeric actin (α-SA) (red). Nuclei were stained with DAPI (blue). Scale bar, 100 µm. i) Quantification results of PT BsAbs in the MI heart 6 h after inhalation or i.v. injection. N = 6.

  • conjugation
    conjugation

Protocols

  • Protocol Booklet - Chinese Version
  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab201802? Please let us know so that we can cite the reference in this datasheet.

ab201802 has been referenced in 4 publications.

  • Boonpiyathad T  et al. Allergen-specific immunotherapy boosts allergen-specific IgD production in house dust mite-sensitized asthmatic patients. Allergy 75:1457-1460 (2020). PubMed: 31769883
  • Liu M  et al. Bispecific Antibody Inhalation Therapy for Redirecting Stem Cells from the Lungs to Repair Heart Injury. Adv Sci (Weinh) 8:2002127 (2020). PubMed: 33437573
  • Wu X  et al. Ficolin A derived from local macrophages and neutrophils protects against lipopolysaccharide-induced acute lung injury by activating complement. Immunol Cell Biol N/A:N/A (2020). PubMed: 32339310
  • Boonpiyathad T  et al. Role of Der p 1-specific B cells in immune tolerance during 2 years of house dust mite-specific immunotherapy. J Allergy Clin Immunol 143:1077-1086.e10 (2019). PubMed: 30529452

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