DyLight® 488 Conjugation Kit (Fast) - Lightning-Link®

• Flow Cyt

PubMed

Flow Cytometry - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

Monaco, Sara, et al used DyLight^® 488 Conjugation Kit (Fast) - Lightning-Link^® (ab201799) as part of examining the co-localization of Smoothened and AC3 in primary cilia. They used the kit to conjugate DyLight^® 488 to anti-AC3 antibody for use in flow cytometry.
(A) Representative dot plots of untreated (Control) and SAG-treated samples (SAG) from E18 embryos immunostained for AC3-FITC and Smoothened-APC (Smo-APC). (B) Quantification of the percentage of Smo+AC3+ particles. Bar graphs show mean ± SEM, p-values are calculated with Student's t-test. Statistically significant differences are indicated with asterisks (*p < 0.05, **p < 0.01).

Image from Monaco, Sara, et al., Front Cell Neurosci., 12:519; doi: 10.3389/fncel.2018.00519. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Schematic Diagram

Supplier Data

Schematic Diagram - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

This illustration demonstrates a general procedure of how the Lightning-Link®(Fast) labeling technology enables the direct labeling of antibodies or proteins.

Simply pipette your antibody or biomolecule of choice into the vial of a lyophilized mixture containing the label of interest and incubate for just 15 minutes. Please see the ab201799 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

Lalo, Ulyana, et al used DyLight^® 488 Conjugation Kit (Fast) - Lightning-Link^® (ab201799) as part of examining the colocalization of vesicular ATP transporters with exocytotic organelle markers. They used the kit to conjugate DyLight^® 488 to various antibodies for use in multiphoton fluorescent microscopy.
Living, acutely isolated cortical astrocytes were labeled with antibodies to vesicular transporters (VNUT1 and VGLUT1) and synaptic vesicle (SV2A) and lysosomal (LAMP3 and Cathepsin-D) markers. Antibodies were conjugated to fluorescent dyes DyLight488 (green) and DyLight594 (red) prior to astrocyte labeling. (A-D) The representative two-photon fluorescence images (maximal intensity projections of Z-stack) and results of colocalization analysis, carried out using NIH ImageJ 1.43 software. The correlation between green and red fluorescence (images in the right column) is depicted as a product of the relative differences from the mean (PDM) for each pixel; the pseudocolor PDM images were generated as an output of ImageJ analysis routine. Positive values (bright yellow) are indicative for good co-localization of green and red signals, negative values (blue-violet) indicate segregation, and black color shows the lack of correlation. Note the different extent of scale for PDM values in (A-D). All scale bars in (A-D) are 5 μm.

Image from Lalo, Ulyana, et al., PLoS Biol., 12(1): e1001747; doi: 10.1371/journal.pbio.1001747. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

Dorfmueller, Simone, et al used DyLight^® 488 Conjugation Kit (Fast) - Lightning-Link^® (ab201799) as part of characterizing antibody-labelled human corneal endothelial cells (hCECs) and stromal fibroblasts. They used the kit to conjugate DyLight^® 488 to C1M2-2-B8 scFv or IgG-B8 full-size antibody for use in flow cytometry.
Primary hCECs or stromal fibroblasts were incubated with C1M2-2-B8 scFv or IgG-B8 full-size antibody conjugated to DyLight^® 488 (4 μg/ml). (a) The histogram for unlabelled hCECs (black) is overlaid with those for scFv and IgG-B8 labelled cells as indicated. Similarly, the histogram for unlabelled stromal fibroblasts (orange) is overlaid with those for scFv and IgG-B8 labelled fibroblasts. (b) The histograms for C1M2-2-B8 scFv labelled hCECs and stromal fibroblasts were shown on the same graph, as for the histograms for IgG-B8 labelled hCECs and stromal fibroblasts. The peak separation between fibroblasts and hCECs were similar in magnitude with either scFv or IgG labelling.

Image from Dorfmueller, Simone, et al., Sci rep., 6:21661; doi: 10.1038/srep21661. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

Mellema, M., et al used DyLight^® 488 Conjugation Kit (Fast) - Lightning-Link^® (ab201799) as part of examining DyLight^® 488-conjugated alendronate binding of calcifying nanoparticles (CNP) generated in vitro. They used the kit to conjugate DyLight^® 488 to alendronate (that selectively and specifically bind to calcium-phosphorus surfaces) for use in Nanoparticle Tracking Analysis (NTA^®).
CNP in the whole urine samples and in vitro generated CNP suspensions were analyzed using the NanoSight LM10HS-48814TS instrument (Malvern Instruments, Worcestershire, UK) with a 488 nm wavelength laser, fluorescent filter (505 nm long pass), and both temperature control and syringe pump modules. A representative histogram shows averaged sizing and relative abundance data (left; SE in red) and individual analyses from the assessment (right) of this representative sample in triplicate. Note that CNP generated in vitro do not acquire the protein/mucus coating known to occur in vivo and the larger forms retain the ability to bind to alendronate.

Image from Mellema et al., PLoS One, 11(12):e0166045; doi: 10.1371/journal.pone.0166045. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - DyLight® 488 Conjugation Kit (Fast) - Lightning-Link® (AB201799)

DyLight® 488 Conjugation Kit - Lightning-Link® labeling alendronate compound and osteopontin protein for Nanoparticle Tracking Analysis (NTA®).

Mellema M et al. used ab201799 as part of examining mineralo-organic nanoparticles in feline urine.

They used the kit to conjugate DyLight^® 488 to alendronate compound and osteopontin protein for use in Nanoparticle Tracking Analysis (NTA^®).

A : Representative histogram showing size distribution and cumulative percentage (left) of submicron particulate matter in healthy feline urine after labeling with DyLight 488 conjugated alendronate. A representative frame from the captured video analyzed by NTA is shown as well (right). Note the strongly preferential binding to primary CNP that lack an outer layer of protein or mucus.

B : Representative histogram showing averaged sizing and relative abundance data (left; SE in red) and individual analyses from the assessment (right) of this representative sample of healthy feline urine after labeling with DyLight 488 conjugated osteopontin. Note the strongly preferential binding to primary naturally-occurring CNP that lack an outer layer of protein or mucus.

Image from Mellema M et al., PloS One,11(12):e0166045. Fig 4.; doi: 10.1371/journal.pone.0166045. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/