PE/Cy7® Conjugation Kit - Lightning-Link®

• Conjugation

PubMed

Conjugation - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

Oliveira BM et al. used ab102903 PE-Cy7^® conjugation kit with a mouse anti-bovine CD45RO antibody and ab102859 APC-Cy7^® conjugation kit with a mouse anti-Bovine CD62L antibody. This enabled them to run their desired multicolor flow cytometry panel.

Data shows flow cytometry gating strategy used to define γδ T cells (TCRγδ+CD3+CD335−), CD4+ T cells (CD4+CD3+TCRγδ−CD335−), CD8+ T cells (CD8+CD3+TCRγδ−CD335−) and NK cells (CD335+CD3−) in the stromal vascular fraction (SVF) of mesenteric and subcutaneous bovine adipose tissue (MAT and SAT, respectively) and in peripheral blood leukocytes. Dead cells were excluded with Fixable Viability Dye (FVD), lymphocytes were gated based on SSC-A versus FSC-A and singlets were selected from the FSC-A versus FSC-H dot plot.

The flow cytometry gating strategy used to define CD45RO+ and CD62L+ T cell subpopulations is also shown in CD8+

Image from Oliveira BM et al., Scientific reports., 9 (1) 3413. Fig 1.; doi: 10.1038/s41598-019-39938-0. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• Schematic Diagram

Supplier Data

Schematic Diagram - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

This illustration demonstrates a general procedure of how Lightning-Link® 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 around 3 hours. Please see the ab102903 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

Flores, Erica B et al. used PE/Cy7^® Conjugation Kit - Lightning-Link^® as part of examining Poxiviruses. They used the kit to conjugate sulfated heparin (clone F58-10E4) for use in flow cytometry.
Sulfation of cell surface heparin measured via flow cytometry utilizing an antibody that recognizes the sulfated 10E4 epitope on HS chains. The mean fluorescent intensity (MFI) values were then normalized to NDST+ values to show the relative change in the NDST-/- cell lines

Image from Flores et al., PloS one, 15(4): e0231977; doi: 10.1371/journal.pone.0231977. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

Nishimori, Asami et al used PE/Cy7^® Conjugation Kit - Lightning-Link^® (ab102903) as part of examining bovine leukemia virus infection. They used the kit to conjugate PE/Cy7^® to anti-bovine IgM antibody for use in flow cytometry.
A BLV-infected cow (#368, Holstein, female, 538 kg, 31 months old) was inoculated with 530 mg (1 mg/kg) of the purified 4G12 intravenously. (A) The proliferation of CD4+ and CD8+ T cells against BLV antigen. Peripheral blood mononuclear cells (PBMCs) isolated from the cow which was inoculated with 4G12 were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and cultured without stimulation (medium) or with the supernatant of FLK or FLK-BLV cells for 6 days. After the cultivation, the proliferation of T cells was immediately analyzed by flow cytometry. A P-value less than 0.05 was considered statistically significant. #, P <0.05 (FLK-BLV, versus day 0; one-way ANOVA followed by Dunnett's test). (B) Changes in PD-L1 occupancy on circulating IgM+ B cells calculated by the binding of 4G12 to bovine PD-L1. The occupancy was estimated as the percentage of the in vivo PD-L1 binding occurred at the total available binding sites. (C) Changes in BLV provirus loads in the cow inoculated with 4G12; the y-axis shows the number of BLV copies included in 50-ng DNA extracts of PBMCs. Data are means ± SEM of at least three replicate experiments.

Image from Nishimori, Asami et al. PloS one vol. 12,4 e0174916. 26 Apr. 2017, doi:10.1371/journal.pone.0174916. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

Flow Cytometry - PE/Cy7 Conjugation Kit;- Lightning-Link.

Okagawa, Tomohiro, et al used PE/Cy7^® Conjugation Kit - Lightning-Link^® (ab102903) as part of examining PD-1 and LAG-3 expression. They used the kit to conjugate PE/Cy7^® to monoclonal anti-IgM antibody, clone IL-A30, for use in flow cytometry.
Expression of PD-1 and LAG-3 on CD4+ T cells in BLV-infected cattle. A Gating strategy and representative dot plots for expression analyses of PD-1 and LAG-3 on IgM−CD3+CD4+γδTCR− T cells from peripheral blood of BLV-infected cattle (AL and EBL). Values in the quadrants indicate percentages of cells. Percentages of PD-1+LAG-3+CD4+ T cells (B), PD-1+LAG-3−CD4+ T cells (C), and PD-1−LAG-3+CD4+ T cells (D) in CD3+CD4+ T-cell population in peripheral blood from BLV-uninfected (BLV − ; n = 15), AL (n = 22), PL (n = 11), and EBL cattle (n = 7). Bars indicate group median percentage. Significant differences between each group were determined using a Kruskal–Wallis test, where P < 0.05 and P < 0.001, indicated by asterisks (* and ***, respectively).

Image from Okagawa, Tomohiro et al., Vet Res., 49(1):50. doi: 10.1186/s13567-018-0543-9. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (AB102903)

Flow Cytometry - PE/Cy7 Conjugation Kit- Lightning-Link.

Robinson, Andrew P., et al used PE/Cy7^® Conjugation Kit - Lightning-Link^® (ab102903) as part of characterizing oligodendroglial populations. They used the kit to conjugate PE/Cy7^® to Mouse anti-A2B5 antibody, clone 105, for use in flow cytometry.
SJL/J mice were immunized with PLP139–151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n = 5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45− or CD45low. (B) Oligodendroglial cells were defined by double positive staining : A2B5+PDGFRα+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes.

Image from Robinson, Andrew P., et al., PloS one, 9(9):e107649. doi: 10.1371/journal.pone.0107649. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/