Multiplex IHC staining was performed to examine the co-expression of ATAD2 and Ki-67 in OC tissue sections on a commercially obtained OC tissue microarray (OV241c; containing 12 primary OC tissues) from Alenabio using an Opal™ 4-Color Manual IHC kit (PerkinElmer, Inc.). Briefly, the sections were first incubated with a specific ATAD2 antibody (1:1,000; catalog no. HPA029424; Sigma-Aldrich; Merck KGaA) for 1 h at room temperature, followed by incubation with a goat anti-rabbit immunoglobulin G antibody (1:5,000; catalog no. NEL840001KT, PerkinElmer, Inc.) for 1 h at room temperature and subsequently labeled with Opal 520 fluorophore (PerkinElmer, Inc.). For Ki-67 staining, an anti-Ki67 antibody (1:1,000; catalog no. ZM-0166; OriGene Technologies, Inc.) was used at room temperature for 1 h and Opal 690 fluorophore (PerkinElmer, Inc.) was used for labeling. Finally, nuclei were stained with DAPI (PerkinElmer, Inc.) for 30 min at room temperature and mounted with anti-fade medium. The inForm 2.4.2 software (PerkinElmer, Inc.) was used to analyze the multispectral images.
Opal 520 fluorophore
Manufactured by PerkinElmer
The Opal 520 fluorophore is a dye molecule designed to emit fluorescent light when exposed to specific wavelengths of light. It is a core component used in various analytical instruments and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Opal 690 fluorophore, by PerkinElmer (2 mentions)
Opal 4 color manual ihc kit, by PerkinElmer (2 mentions)
Inform 2, by PerkinElmer (1 mentions)
Hpa029424, by Merck Group (1 mentions)
Zm 0166, by OriGene (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by PerkinElmer (1 mentions)
Inform software, by PerkinElmer (1 mentions)
Opal polymer hrp ms rb, by PerkinElmer (1 mentions)
2 protocols using opal 520 fluorophore
1
Multiplex IHC for ATAD2 and Ki-67 in OC
2
Multiplexed Immunohistochemistry of Tn Antigen and MUC2 in CRC
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Totally, 48 formalin‐fixed paraffin‐embedded sections of human CRC tissues were stained with Tn antigen and mucin MUC2 simultaneously performed with OpalTM 4‐Color Manual IHC Kit (PerkinElmer). Briefly, the sections were first incubated with a specific anti‐Tn IgM mAb(CA3638, clone 12A8‐C7‐F5) for 1 hour at room temperature and then incubated with Goat antimouse IgM mu chain (HRP)(Abcam, ab97230) and eventually labelled with Opal 520 fluorophore (PerkinElmer). For the staining of MUC2, Goat anti‐MUC2 antibody (2 μg/mL, Santa Cruz, sc‐3739) was applied and Opal polymer HRP Ms+Rb (PerkinElmer) was subsequently added to the sections followed by incubation with Opal 690 fluorophore (PerkinElmer). Finally, all sections were counterstained with DAPI and mounted with anti‐fade mountants. The inForm software (PerkinElmer) was used for analysing the multispectral images.
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