DyLight® 594 Conjugation Kit (Fast) - Lightning-Link®

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (AB201801)

Lalo, Ulyana, et al used DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (ab201801) as part of examining the colocalization of vesicular ATP transporters with exocytotic organelle markers. They used the kit to conjugate DyLight® 594 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/

• Schematic Diagram

Supplier Data

Schematic Diagram - DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (AB201801)

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 ab201801 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Flow Cyt

PubMed

Flow Cytometry - DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (AB201801)

Hsiao, Feng, et al used DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (ab201801) as part of examining SAMHD1 expression in tissue Tm subsets. They used the kit to conjugate DyLight® 594 to anti-SAMHD1 antibody for use in flow cytometry.
A) Validation of SAMHD1 antibody by flow cytometry. Unstimulated PBMCs were mock-treated or exposed for 3 days to F4.HSA. Depicted within the gates are the proportions of infected cells amongst the SAMHD1- and SAMHD1+ cells. The results demonstrate that SAMHD1- cells, which are permissive, harbor a higher proportion of infected cells than non-permissive, SAMHD1+ cells, as expected [3]. Results are pre-gated on live, singlet CD3+CD8-CD45RO+CD45RA- cells. B) Bar graph of the percentage of SAMHD1+ cells across the three indicated memory CD4+ T cell subsets from HLACs as determined by flow cytometry. Results are pre-gated on live, singlet CD3+CD8-CD45RO+CD45RA- cells. ns : non-significant as determined using a 2-tailed paired parametric t-test.

Image from Hsiao, Feng, et al., PLoS pathog., 16(4): e1008450; doi: 10.1371/journal.ppat.1008450. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/