PE/Cy5® Conjugation Kit - Lightning-Link®

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

Bardhan, Kankana, et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) as part of examining PD-1pY248+ (pPD-1) expression in human T cells. They used the kit to conjugate PE/Cy5® to Anti-PD-1pY248 antibody for use in flow cytometry.
pPD-1 is predominantly expressed in CD8+ TCM cells. CCR7 and CD45RO markers were used to identify central memory (TCM) and effector memory (TEM) T cells. After gating on CD45RO expression, TCM (CD45RO+CCR7+) and TEM (CD45RO+CCR7) CD4+ and CD8+ T cells were identified by assessing expression of CCR7. In TCM and TEM populations, expression of PD-1 was determined and, subsequently, expression of pPD-1 (pPD-1-Y248) was assessed in the PD-1+ population within each subset. Results are representative of six separate experiments.

Image from Bardhan, Kankana, et al., Scientific reports, 9(1):17252. doi: 10.1038/s41598-019-53463-0 .Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Schematic Diagram

Supplier Data

Schematic Diagram - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

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 ab102893 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

Kim, Nayoung, et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) as part of examining apoptosis in HIV-1-infected CD4+ primary T cells. They used the kit to conjugate PE/Cy5® to anti-FOXO3a 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/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

Nielsen, Carolyn M., et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) to conjugate PE/Cy5® to anti-IL-12Rβ2 antibody for use in flow cytometry.
NK cells were analyzed for surface expression of 12Rβ2 using an mIgG1 PECy5-conjugated isotype control to set the gate.

Image from Nielsen, Carolyn M., et al., J Immunol., 194(10):4657-67; doi: 10.4049/jimmunol.1403080. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/3.0/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

Olling, Alexandra, et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) as part of examining the role of toxins in the pathogenicity of Clostridium difficile. They used the kit to conjugate PE/Cy5® to for use in flow cytometry.Binding of fluorescent labeled TcdA-PE/Cy5 and TcdA11874-Atto488 to HT29 and CHO-C6 cells was investigated by FACS analysis. Right shift of the black curve illustrates toxin binding which was detected through fluorescence emission at 667 nm for TcdA and at 523 nm for TcdA1-1874, respectively. Due to different ratio of fluorophor and toxin, fluorescence intensity of TcdA-PE/Cy5 cannot directly be compared with TcdA1-1874-Atto488.

Image from Olling, Alexandra, et al., PLoS One, 6(3):e17623. doi: 10.1371/journal.pone.0017623. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

Taichman, Russell S., et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) as part of examining Axl and Tyro3 expression during experimental prostate cancer (PCa) progression. They used the kit to conjugate PE/Cy5® 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 - PE/Cy5® Conjugation Kit - Lightning-Link® (AB102893)

White, Matthew J., et al used PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893) as part of examining Interleukin‐12 receptor β2 (IL‐12Rβ2) expression relatively to CD57 expression in Peripheral blood mononuclear cells (PBMC). They used the kit to conjugate PE/Cy5® to anti-IL‐12Rβ2 monoclonal antibody for use in flow cytometry.
PBMC were analysed ex vivo for IL‐12Rβ2 expression. (a) Representative flow cytometry plots for IL‐12Rβ2. Frequency (b) and mean fluorescence intensity (MFI) (c) of IL‐12Rβ2 expression were assessed by subset. Each data point represents one donor, n = 19. Lines indicate median values. CD56dim subsets were analysed for linear trend with a repeated measures analysis of variance. ****P ≤ 0.0001.

Image from White, Matthew J., et al., Immunology, 142(1); doi: 10.1111/imm.12239. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/3.0/