DAPI(hydrochloride)

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DAPI(hydrochloride)

本产品不向个人销售，仅用作科学研究，不用于任何人体实验及非科研性质的动物实验。

CAS NO:	28718-90-3
规格:	98%
分子量:	350.3

包装与价格：

包装	价格(元)
5mg	询价
10mg	询价
25mg	询价

DAPI dihydrochloride 是结合富含 A-T 碱基对 DNA 序列的荧光染色。

CAS：28718-90-3
分子式：C16H15N5.2HCl
分子量：350.3
纯度：98%
存储：Store at -20°C

Background:

DAPI, as a DNA-specific probe, can form a fluorescent complex by attaching in the minor grove of A-T rich sequences of DNA and nonfluorescent intercalative complexes with double-stranded nucleic acids. DAPI, as a chromosome and nuclear stain, has maximum excitation ultraviolet (UV) light wavelength with 358 nm and emission in the blue range with 461 nm.[1]

DAPI is usually used for flow cytometry, chromosome staining, DNA visualization and quantitation in histochemistry and biochemistry.[2] DAPI is a good substrate of hOCT1 with a Michaelis constant of 8.94 μM.[6].

In vitro, DAPI (1.43μM) can measure mitochondrial permeability transition and mitochondrial membrane depolarization by combining with Annexin V(FITC)and the potentiometric fluorescent dye, tetramethylrhodamine methyl ester (TMRM).[3] In vitro, to evaluate the DAPI fluorescence, SSC buffer (pH 7.2), containing 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04% SDS or Triton X-100 and 2 μg/mL of DAPI dye, there was found that a linear relationship between DAPI fluorescence and Triton X-100 concentrations ranging from 0.01 to 0.04%.[4] In addition, when the concentration of DAPI was increased from 0.4 μM to 400 μM, the total signal intensity increased, implying an increase in the concentration of DAPI molecules bound to the chromosomes.[5] In vitro experiment it shown that treatment with NR 1 μg/mL, DAPI 5 μg/mL, ORO 0.3% (v/v) and CV 0.5% (v/v) provided optimal linearity and coverage of signals over a range of cell densities (corresponding to 10-100% cell confluence).[7].

参考文献:
[1] Karg TJ, et al. Photoconversion of DAPI and Hoechst dyes to green and red-emitting forms after exposure to UV excitation. Chromosoma. 2018 Jun;127(2):235-245.
[2] Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.
[3] Wallberg F, et al. Analysis of Apoptosis and Necroptosis by Fluorescence-Activated Cell Sorting. Cold Spring Harb Protoc. 2016 Apr 1;2016(4):pdb.prot087387.
[4] Simoliūnas E, et al. DNA-DAPI Interaction-Based Method for Cell Proliferation Rate Evaluation in 3D Structures. Curr Issues Mol Biol. 2021 May 30;43(1):251-263.
[5] Estandarte AK, et al. The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes. Sci Rep. 2016 Aug 16;6:31417.
[6] Yasujima T, et al. Characterization of human OCT1-mediated transport of DAPI as a fluorescent probe substrate. J Pharm Sci. 2011 Sep;100(9):4006-12.
[7] Phan MAT, et al. Semi-quantification of lipids in human meibomian gland epithelial cells using dual staining microplate assays. Exp Eye Res. 2021 Sep;210:108719.

Protocol:

Cell experiment [1]:
Cell lines	bone marrow-derived adherent cells
Preparation Method	After three passages, bone marrow-derived adherent cells were incubated with 50 μg/ml DAPI (Molecular Probes, Invitrogen, Carlsbad, CA) at 37°C and 5% CO2 for 2 h. BM-MSCs were treated with trypsin (Gibco) for 3 min to generate a single-cell suspension, washed three times with DMEM, resuspended in serum-free DMEM, counted, and stored on ice until transplantation. Fluorescence from DAPI labeling on BM-MSCs and cell viability were evaluated for 1, 2, 3, 4, and 8 weeks after cell labeling.
Reaction Conditions	50 μg/ml; at 37°C and 5% CO2 for 2 h
Applications	After three passages in culture, the isolated cell population became homogeneous, showing a monolayer consisting of adherent cells displaying further traits of MSCs, including a typical fibroblast-like morphology and increased proliferation.
Animal experiment [2]:
Animal models	Brain tumor xenograft in nude mice
Preparation Method	Tissue sections for in vivo analysis were collected from animals 3 days after PBS, PAM-RG4/pJDK, or PAM-RG4/pJDK-apoptin injection and immediately paraffinized before further analysis using the DeadEnd(TM) Fluorometric TUNEL System. Following the given protocol, samples were analyzed using fluorescence microscopy to detect the localized green fluorescence of apoptotic cells and blue fluorescence (DAPI) of cell nuclei.
Dosage form	3 days after injection; 0.5-10μg/ml
Applications	Apoptosis, chromatin condensation, which is caused by the degradation of nuclear membrane proteins, by using DAPI staining. however, when applied in vivo, the experimental group did not show any signs of side effects, indicating a sufficiently low level of cytotoxicity, although it showed similar DAPI and TUNEL assay results.
References: [1].Castanheira P, et al. DAPI diffusion after intravitreal injection of mesenchymal stem cells in the injured retina of rats. Cell Transplant. 2009;18(4):423-31. [2].An S, et al. Nonviral gene therapy in vivo with PAM-RG4/apoptin as a potential brain tumor therapeutic. Int J Nanomedicine. 2013;8:821-34.