DyLight® 550 Conjugation Kit (Fast) - Lightning-Link® (ab201800)
Overview
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Product name
DyLight® 550 Conjugation Kit (Fast) - Lightning-Link® -
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.
This product is manufactured by Expedeon, an Abcam company. See the table below to match Abcam kit size against Expedeon product code.
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.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
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Notes
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid DyLight® 550 Labeling Kit. 323-0015 is the same as the 1 mg size. 323-0010 is the same as the 3 x 100 ug size. 323-0030 is the same as the 3 x 10 ug size. 323-0005 is the same as the 100 ug size.
Amount and volume of antibody for conjugation to DyLight® 550
Kit size Recommended
amount of antibody1Maximum
amount of antibodyMaximum antibody
volume23 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 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2mg/ml or
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.
DyLight® is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg ab274016 - Dylight® 550 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
Images
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Staining for mouse-on-mouse immunofluorescence Image courtesy of an Abreview submitted by Bryan Niedenberger.
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.
See Abreview.
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conjugation
