APC/Cy7® Conjugation Kit - Lightning-Link®

• Flow Cyt

PubMed

Flow Cytometry - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

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

Robinson, Andrew P., et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) as part of characterizing oligodendroglial populations. They used the kit to conjugate APC/Cy7^® to Mouse anti-MOG antibody, clone 8-18C5, 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/

• Schematic Diagram

Supplier Data

Schematic Diagram - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

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 ab102859 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Flow Cyt

PubMed

Flow Cytometry - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

Taichman, Russell S., et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) as part of examining Axl and Tyro3 expression during experimental prostate cancer (PCa) progression. They used the kit to conjugate APC/Cy7^® to antibodies for use in flow cytometry.
Anti-Axl, anti-Tyro3 and anti-Ki67 antibodies were conjugated to the fluorophores APC-Cy7, PE-Cy5, and Atto390 using our Lightning-Link^® Conjugation kits. (A) Experimental model. Human PCa cell lines (PC3Luc, DU145Luc) were implanted s.c. into male SCID mice as a model of a primary (1^°) tumor development, and removed after 1 month. At monthly intervals thereafter human PCa cells were identified by anti-HLA staining; proliferative status (Ki67 staining) and Axl or Tyro3 levels were evaluated by FACS. (B) Percent expression of Ki67 by lineage depleted (Lin-) marrow cells or by primary tumor cells at 1 month. (C-D) Percent expression of Axl or Tyro3 by primary tumor cells established with (C) DU145 or (D) PC3 cells or by DTCs recovered from marrow over time.

Image from Taichman, Russell S., et al., PLoS One, 8(4): e61873; doi: 10.1371/journal.pone.0061873. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

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

Okagawa, Tomohiro, et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) as part of examining the effect on proliferation of bovine leukemia virus (BLV)-specific T cells of the administration of Boch5D2. They used the kit to conjugate APC/Cy7^® to monoclonal anti-TCR1-N24 (δ chain) antibody, clone GB21A, for use in flow cytometry.
T-cell proliferation specific for BLV antigen stimulation. Carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled peripheral blood mononuclear cells were cultured in triplicate with fetal lamb kidney (FLK)-BLV antigen, control FLK antigen (A), or gp51 peptides (0.1 and 1 μg/ml) (B) for 6 days. The percentage of CFSElow cells in CD4+ and CD8+γδTCR− T cells was measured by flow cytometry. CFSElow cells represent cells proliferated during cultivation. Each dot represents the mean of three independent experiments. Significant differences were determined by Dunnett's multiple-comparison test across the time points. *,#,†P < 0.05 versus 0 dpi in each stimulation.

Image from Okagawa, Tomohiro, et al., Front Immunol., 8:650, doi: 10.3389/fimmu.2017.00650. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

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

Kopanke, Jennifer H., et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) to identify CD5+/5-ethynyl-2′-deoxyuridine (EdU)+ cells. They used the kit to conjugate APC/Cy7^® to Mouse Anti-Feline CD5 antibody for use in cell proliferation assay.
Gating tree used to identify CD5+/5-ethynyl-2′-deoxyuridine (EdU)+ cells. Single cells were identified by plotting side scatter area by side scatter height. Cells aligning linearly were counted as single cells. CD5+ cells were identified from single cells by plotting FL8 (the channel recognizing APC-Cy7) by side scatter area. Finally, CD5+ EdU+ cells were identified from CD5+ cells by plotting FL6 (the channel that detects APC) by FL8.

Image from Kopanke, Jennifer H., et al., Scientific reports; 8(1):8168. doi: 10.1038/s41598-018-26558-3. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

Okagawa, Tomohiro, et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) as part of examining PD-1 and LAG-3 expression. They used the kit to conjugate APC/Cy7^® to monoclonal anti-TCR1-N24 (δ chain) antibody, clone GB21A, 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/

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

Kim, Nayoung, et al used APC/Cy7^® Conjugation Kit - Lightning-Link^® (ab102859) as part of examining apoptosis in HIV-1-infected CD4+ primary T cells. They used the kit to conjugate APC/Cy7^® to anti-PPP2R1B antibody for use in confocal microscopy.
Jurkat T cells expressing tTA alone, TatSF2+tTA, or TatSF2G48-R57A +tTA were stained with antibodies against PPP2R1B (first and forth columns of panels, green), pFOXO3a (second column, red), and FOXO3a (forth column, red).

Image from Kim, Nayoung, et al., PLoS Pathog., 6(9): e1001103. doi: 10.1371/journal.ppat.1001103. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - APC/Cy7® Conjugation Kit - Lightning-Link® (AB102859)

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+ T cells.

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/.