Thiazine red

Manufactured by Merck Group

Sourced in United States

Thiazine Red is a synthetic dye compound used as a staining agent in various laboratory applications. It is a water-soluble dye that exhibits reddish-purple coloration. The primary function of Thiazine Red is to stain and visualize specific biological structures or compounds during microscopic analysis and other laboratory procedures.

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Lab products found in correlation

5 protocols using thiazine red

Polyelectrolyte Complexes for Cell Studies

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Poly(allylamine hydrochloride) (PAH, Mw = 17.5 kDa), poly(4-styrene sulfonate) sodium salt (PSS, Mw = 70 kDa), calcium chloride dihydrate, sodium carbonate, sodium chloride, ethylenediaminetetraacetic acid (EDTA), dextran sulfate sodium salt (DS, Mw = 40 kDa), rhodamine B, rhodamine 6G, pyronin B, fluorescein, acridine yellow, acridine orange, thiazine red, and polyvinyl alcohol were purchased from Sigma.
Dulbecco's modified Eagle's medium (DMEM), Dulbecco's modified Eagle's medium F12 (DMEM F12), phosphate-buffered saline (PBS), fetal bovine serum (FBS), trypsin EDTA, bovine serum albumin (BSA), calcein-AM ﬂuorescent dye, hoechst 33258 dye, dimethyl sulfoxide (DMSO) and MTT cell proliferation assay ((3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay) kit were purchased from Thermo Fisher Scientific.
Deionized (DI) water (resistivity 18.2 MΩ cm at 25 °C) from the Direct-Q water puriﬁcation system was used to prepare all solutions.

Kozyreva Z.V., Demina P.A., Sapach A.Y., Terentyeva D.A., Gusliakova O.I., Abramova A.M., Goryacheva I.Y., Trushina D.B., Sukhorukov G.B, & Sindeeva O.A. (2024). Multiple dyes applications for fluorescent convertible polymer capsules as macrophages tracking labels. Heliyon, 10(10), e30680.

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Immunofluorescent Staining of Amyloid-β in Brain

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The brain sections were washed with PBS and incubated in PBS/0.1% Triton (T-PBS) for 30 min at 20 °C while shaking. After incubation, the sections were blocked in T-PBS, 20% horse serum (Gibco Invitrogen) for 30 min at 20 °C while shaking. Following blocking, brain sections were incubated with primary Aβ antibody (Aβ, 1–16 (6E10), Covance, 1:1000) in T-PBS and 0.2% bovine serum albumin (BSA) for 2–3 days at 4 °C. The sections where then washed and incubated with anti-mouse fluorescent Alexa 546 antibody (Invitrogen-Life tech, Vienna, Austria) in T-PBS and 0.2% BSA for 1 hour at 20 °C while shaking. Finally the sections were washed and then mounted onto glass slides and coverslipped with Mowiol^® 4–88 (Roth, Austria). Some sections were stained with Thiazine Red or Resorufin (1.6 μg/ml, Sigma, overnight) to label plaques or CAA respectively. Some sections were counterstained with tyrosine hydroxylase antibodies (Novus, NB300-109; 1:1000, secondary anti-rabbit Alexa 488). Some sections were counterstained with blue DAPI (1:10 000, 1 hour) to visualize nuclei.

Kniewallner K.M., Wenzel D, & Humpel C. (2016). Thiazine Red+ platelet inclusions in Cerebral Blood Vessels are first signs in an Alzheimer’s Disease mouse model. Scientific Reports, 6, 28447.

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Investigating Fibrinolytic Mechanisms in Immune Cells

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Reagents were from Life Technologies unless indicated otherwise. Recombinant human t-PA was Actilyse (Boehringer, Ingelheim, Germany). Human plasminogen, human fibrinogen and bovine thrombin were from Merck Millipore (Kilsyth, Victoria, Australia). Human and mouse plasmin were from Hematologic Technologies (Essex Junction, Vermont, USA). Thiazine Red, staurosporine, 6-aminocaproic acid, lipopolysaccharide (LPS), aprotinin, PKH26 and PKH67 fluorophores were from Sigma-Aldrich (St. Louis, Missouri, USA). Recombinant human/mouse IL-4 (rIL-4) and recombinant human/mouse GM-CSF (rGM-CSF) were from Peprotech (Rocky Hill, New Jersey, USA). Ficoll-Paque was from GE Healthcare (Rydalmere, New South Wales, Australia). [H³]-thymidine was from Amersham (Little Chalfont, Buckinghamshire, U.K.). Protease inhibitor tablets for cell lysis were from Roche (Mannheim, Germany).

Borg R.J., Samson A.L., Au A.E., Scholzen A., Fuchsberger M., Kong Y.Y., Freeman R., Mifsud N.A., Plebanski M, & Medcalf R.L. (2015). Dendritic Cell-Mediated Phagocytosis but Not Immune Activation Is Enhanced by Plasmin. PLoS ONE, 10(7), e0131216.

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Immunofluorescence Analysis of Amyloid-β in Mice

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Mice were deeply anesthetized with a mixture of ketamine (300 mg per kg) and xylazine (20 mg per kg) and transcardially perfused with ice‐cold PBS and 4% paraformaldehyde. Brains were removed and postfixed for 24 h in 4% paraformaldehyde (Roti^®‐Histofix, Roth), followed by 48 h in 30% sucrose (in PBS). Frozen brains were cut into 25‐µm‐thick coronal serial sections on a sliding microtome (SM2000R, Leica Biosystems, Wetzlar, Germany) and collected in 5% Gycerol (in PBS). Immunofluorescence staining was performed using the following antibodies: rabbit polyclonal antibody 3552 specific for Aβ 1‐40 (kindly provided by E. Kremmer, Ludwig Maximilians University, Munich, Germany; diluted 1:3000), rabbit anti‐doublecortin (DCX; 1:5000; abcam), mouse monoclonal anti‐NeuN (1:200; Millipore) and DAPI (Roche) was used as a counterstain. Dense‐core plaques were stained with Thiazine Red (Sigma Aldrich; 2 µM solution in PBS for 5 minutes at RT followed by 3 × 5 minutes washes). Appropriate secondary antibodies conjugated to Alexa 488 or 555 (1:1500) were used.

Katzmarski N., Ziegler‐Waldkirch S., Scheffler N., Witt C., Abou‐Ajram C., Nuscher B., Prinz M., Haass C, & Meyer‐Luehmann M. (2019). Aβ oligomers trigger and accelerate Aβ seeding. Brain Pathology, 30(1), 36-45.

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Immunohistochemistry for Tau and Neurofilament Proteins

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Slices were postfixed and immunohistochemistry was performed as described by us (Kniewallner et al., 2016 (link)). Briefly, brain slices were washed with PBS and incubated in PBS/0.1% Triton (T-PBS) for 30 min at room temperature (RT) while shaking. After incubation, the sections were blocked in T-PBS, 20% horse serum (Gibco Invitrogen) for 30 min at 20°C while shaking. Following blocking, slices were incubated with 5 μl/well sample reducing agent (Invitrogen-Life tech, Vienna, Austria) for 10 min at 70°C cooled on ice and then incubated with the primary antibody tau-5 (1:250, Thermo Fisher, AHB0042), neurofilament-200 kDa (1:1000, Novus) and PHF Tau (p-Tau 202) clone AT8 antibody (1:500, Thermo MN1020) as well as phospho-tau-T231 (1:250, BioLegend, 807201; phospho-tau-S396, 1:250, BioLegend, 807401), in T-PBS and 0.2% bovine serum albumin (BSA) for 2–3 days at 4°C. The sections where then washed and incubated with anti-rabbit fluorescent Alexa 488 antibody (1:400, Invitrogen-Life Tech, Vienna, Austria) in T-PBS and 0.2% BSA for 1 h at 20°C while shaking. Finally the sections were washed and then mounted onto glass slides and coverslipped with Mowiol^® 4-88 (Roth, Austria). Alternatively, sections were stained with Thiazine Red (1.6 μg/ml, Sigma, overnight) to label plaques and counterstained with DAPI (1:10,000, 1 h) to visualize nuclei.

Foidl B.M, & Humpel C. (2018). Differential Hyperphosphorylation of Tau-S199, -T231 and -S396 in Organotypic Brain Slices of Alzheimer Mice. A Model to Study Early Tau Hyperphosphorylation Using Okadaic Acid. Frontiers in Aging Neuroscience, 10, 113.

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