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Pe cy7 conjugation kit

Manufactured by Abcam
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The PE/Cy7 conjugation kit is a product that enables the conjugation of PE (phycoerythrin) and Cy7 fluorophores to proteins or other biomolecules. The kit provides the necessary components and protocols to facilitate this labeling process.

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Lab products found in correlation

Percp cy5.5 conjugation kit percp cy5.5, by Bio-Rad (2 mentions) Apc cy7 conjugation kit, by Abcam (2 mentions) Novocyte advanteon flow cytometer system, by Agilent Technologies (1 mentions) Facsuite software, by BD (1 mentions) Apc conjugation kit, by Abcam (1 mentions) Apoptosis detection kit 1, by BD (1 mentions) Facsuite software v 1, by BD (1 mentions) Cellquest version 5, by BD (1 mentions) Anti cd86, by Abcam (1 mentions) Facscalibur, by BD (1 mentions)

6 protocols using pe cy7 conjugation kit

1

Flow Cytometry Analysis of Plasminogen Receptor

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Cell lines were stained with either anti-Plg-RKT mAB, normal mouse IgG2a or unstained. The anti-Plg-RKT and isotype control antibody were directly labeled with PE-Cy7 Conjugation Kit from Abcam (Boston, MA, USA). The stained cells were acquired in a NovoCyte (ACEA Biosciences, San Diego, CA, USA) and analyzed with FlowJo software (Tree Star Inc., Ashland, OR, USA). Viable cells (propidium iodide and annexin V negative) were gated from nonviable cells.
Quantitative flow cytometric equilibrium binding of fluorescein isothiocyanate (FITC)-plasminogen to cells was analyzed using beads impregnated with FITC as described [19 (link)]. Nonspecific (FITC)-plasminogen binding in the presence of epsilon amino caproic acid (EACA) was subtracted from total binding to obtain specific binding for Scatchard analysis.
Miles L.A., Krajewski S., Baik N., Parmer R.J, & Mueller B.M. (2022). Plg-RKT Expression in Human Breast Cancer Tissues. Biomolecules, 12(4), 503.
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2

CRT Expression Analysis in HCT116 and A549 Cells

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HCT116 (1−1.5 × 105) cells and A549 cells (1−1.5 × 105) were harvested and washed twice with PBS. The cells were then incubated for 1.5 h in the dark at 4 °C with PE-Cy-7-labeled anti-CRT that had been labeled with the Abcam PE/Cy7 conjugation kit following the manufacturer’s instructions. HCT116 and A549 cells were then re-suspended in cold PBS (1.5 mL) for analysis on the NovoCyte Advanteon flow cytometer system (Agilent Technologies, Santa Carla, US). Natural cell fluorescence was monitored by excitation at 564 nm and emission at 606 nm; whereas CRT fluorescence was monitored by excitation at 496 nm and emission at 774 nm. Results from the flow cytometer were analyzed with BD FACSuite Software (BD Biosciences, San Jose, CA).
Blair I., Fan J., Gillespie K, & Mesaros C. (2024). Ferroptosis and HMGB2 induced calreticulin translocation required for immunogenic cell death are controlled by the nuclear exporter XPO1. Research Square.
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3

Comprehensive Bovine Immune Cell Profiling

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To construct a seven-colour panel, antibodies were first labelled with conjugation kits, accordingly to manufacturer instructions, as commercially available antibodies for bovines are conjugated to a very limited number of fluorochromes. Namely, mouse anti-bovine CD11b (clone CC126, Bio-Rad) was conjugated to Phycoerythrin (PE) with PE conjugation kit (PE) (AbD Serotec), mouse anti-bovine CD14 (Clone CC-G33, Bio-Rad) was conjugated to peridinin-chlorophyll protein-cychrome 5.5 (PerCP-Cy5.5) with PerCP-Cy5.5 conjugation kit (PerCP-Cy5.5) (Bio-Rad), mouse anti-bovine MHC-II (clone CAT82A, Washington State University) was conjugated to PE-cychrome 7 (PE-Cy7) with PE-Cy7® conjugation kit (Abcam, Cambridge, UK), mouse anti-bovine CD11c (clone BAQ153A, Washington S. U.) was conjugated to Allophycocyanin (APC) with APC conjugation kit (Abcam) and mAb clone CH138A (Washington State University) described to bind to granulocytes, was conjugated to allophycocyanin Cyanine 7 (APC-Cy7) with APC-Cy7® conjugation kit (Abcam). Fluorescein isothiocyanate (FITC) anti-bovine CD45 (Clone CC1, Bio-Rad, Kidlington, UK) was commercially available. All the antibodies used in this study were titrated with the optimal concentration determined for adipose tissue samples.
Oliveira B.M., Pinto A., Correia A., Ferreira P.G., Vilanova M, & Teixeira L. (2020). Characterization of Myeloid Cellular Populations in Mesenteric and Subcutaneous Adipose Tissue of Holstein-Friesian Cows. Scientific Reports, 10, 1771.
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4

Calreticulin Cell Surface Expression and Apoptosis Analysis

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Flow cytometry was used to detect calreticulin on the cell surface of HCT116 and RKO and their derivative clones mir27a_KD or mir27a_OE in basal condition, upon MTX or OXP treatment. Briefly, cells were plated, harvested and washed twice with PBS and incubated for 1 h in darkness at 4 °C with a custom PE-Cy-7-labeled anti-CRT, using PE/Cy7 conjugation kit (ab102903; Abcam), as manufacturer's protocol. Cells were then washed and resuspended in cold PBS for FACS analysis. Apoptosis was evaluated by AnnexinV-PE, 7-AAD using the Apoptosis Detection Kit I (BD Biosciences), according to the manufacturer's protocols. All flow cytometry results were analyzed with FACSuite Software v.1.0.5.3841 (BD Biosciences).
Colangelo T., Polcaro G., Ziccardi P., Muccillo L., Galgani M., Pucci B., Rita Milone M., Budillon A., Santopaolo M., Mazzoccoli G., Matarese G., Sabatino L, & Colantuoni V. (2016). The miR-27a-calreticulin axis affects drug-induced immunogenic cell death in human colorectal cancer cells. Cell Death & Disease, 7(2), e2108-.
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5

Quantifying M1 Phenotype Kupffer Cells

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To determine the ratio of M1 phenotype KCs, 1x106 KCs were suspended in cell staining buffer before incubation with anti-CD68 (cat. no. ab201340; Abcam) and anti-CD86 (cat. no. ab213044; Abcam) antibodies at 4°C for 0.5 h. After washing 3 times with cell staining buffer, the samples were incubated with a FITC conjugation kit (Fast) (cat. no. ab188285; Abcam) or PE/Cy7 conjugation kit (cat. no. ab102903; Abcam) at 4°C in the dark for 30 min. After washing 3 times with cell staining buffer, the stained cells were acquired by a BD FACSCalibur and analyzed by using Cell Quest version 5.1 software (BD Biosciences).
Wang J., Deng M., Wu H., Wang M., Gong J., Bai H., Wu Y., Pan J., Chen Y, & Li S. (2020). Suberoylanilide hydroxamic acid alleviates orthotopic liver transplantation-induced hepatic ischemia-reperfusion injury by regulating the AKT/GSK3β/NF-κB and AKT/mTOR pathways in rat Kupffer cells. International Journal of Molecular Medicine, 45(6), 1875-1887.
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6

Multicolor Flow Cytometry Panel for Bovine Cells

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As commercially available antibodies for bovines are conjugated to a very limited number of fluorochromes, to construct an eight-colour multicolour panel, some of the antibodies were first labelled with conjugation kits accordingly to manufacture instructions. Namely, mouse anti-bovine TCRγδ (Clone GB21A, Washington State University, Pullman, WA) was conjugated to Fluorescein isothiocyanate (FITC) with FITC conjugation kit (Bio-Rad, Kidlington, UK), mouse anti-bovine CD335 (clone AKS1, Bio-Rad) was conjugated to peridinin-chlorophyll protein-cychrome 5.5 (PerCP-Cy5.5) with PerCP-Cy5.5 conjugation kit (PerCP-Cy5.5) (Bio-Rad), mouse anti-bovine CD45RO (clone IL-A116, Bio-Rad) was conjugated to PE-cychrome 7 (PE-Cy7) with PE-Cy7® conjugation kit (Abcam, Cambridge, UK), mouse anti-Bovine CD62L (clone BAQ92A, Washington State University) was conjugated to allophycocyanin Cyanine 7 (APC-Cy7) with APC-Cy7® conjugation kit (Abcam) and mouse anti-bovine CD3 (Clone MM1A, Washington State University) was conjugated to CF™405 M with Mix-n-Stain™CF™405 M Antibody Labeling Kits (Sigma-Aldrich). Phycoerythrin (PE) anti-bovine CD8α (clone CC63) and Alexa Fluor 647® anti-bovine CD4 (clone CC8) (all Bio-Rad) were commercially available. All antibodies were titrated and optimal concentration for adipose tissue samples determined.
Oliveira B.M., Rasteiro A.M., Correia A., Pinto A., Meireles P., Ferreira P.G., Vilanova M, & Teixeira L. (2019). T cells in mesenteric and subcutaneous adipose tissue of Holstein-Friesian cows. Scientific Reports, 9, 3413.
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