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> Apocynin [498-02-2]

Apocynin [498-02-2]

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Cat# HY-N0088-1ml

Size : 10mM/1mL

Brand : MedChemExpress

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Description

Apocynin is a selective NADPH-oxidase inhibitor with an IC50 of 10 μM[1][2]. Apocynin improves acute lung inflammation in Carrageenan (HY-125474)-induced pleurisy mice model[3]. Apocynin can also be used for cancer research[4]. Apocynin reverses the aging process in mesenchymal stem cells to promote osteogenesis and increases bone mass[5].

Cellular Effect
Cell Line Type Value Description References
Neutrophil IC50
3.9 μM
Compound: Apocynin
Antioxidant activity in human neutrophils assessed as inhibition of oxidative burst-induced ROS production by chemiluminescence assay
Antioxidant activity in human neutrophils assessed as inhibition of oxidative burst-induced ROS production by chemiluminescence assay
[PMID: 24588269]
In Vitro

Apocynin (100 μM; 1-7, 14 days) shows a significant increase in the expression level of an osteogenic marker in the aging BMSCs after osteogenic induction[3].
Apocynin (1, 10, 100 μM, 0-48h ) has selective inhibition the proliferation and adhesion to fibronectin of v-H-ras-transformed 3Y1 cells[4].
Apocynin (1, 10, 100 μM; 3, 6, 12 h) decreases the intracellular reactive oxygen species (ROS) level in HR-3Y1-2 but not 3Y1 cells. [4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Apocynin Related Antibodies

Western Blot Analysis[3]

Cell Line: Bone marrow stromal cells (BMSCs)
Concentration: 100 μM
Incubation Time: 1-7,14 days
Result: Increased the expression levels of the “stemness markers” Nanog and Oct-4.
Decreased the expression levels of p53, p21 and p16 at both the mRNA level and protein level.
Increased the expression of sox-2 and klf-4 by 82.4% and 38.7%, respectively, compared with the negative control group.
Reduced the expression of NADPH oxidase by 66.5% compared with the negative control.
Had no change in the cell cycle or proliferation.
Ddecreased the percentage of SA-β –gal-positive (green-stained) cells was by 42.5%.
Increased the expression levels of four pivotal osteogenic markers (Runx2, OSX, Ocn, and Col1).

Cell Proliferation Assay[4]

Cell Line: HR-3Y1-2, 3Y1 cells
Concentration: 0, 1, 10, or 100 μM
Incubation Time: 48h
Result: Inhibited the proliferation of HR-3Y1-2 but not 3Y1 cells at 10μM and 100μM.

RT-PCR[3]

Cell Line: HR-3Y1-2, 3Y1 cells
Concentration: 0, 1, 10, or 100 μM
Incubation Time: 24, 36, 48 h
Result: Selectively down-regulated 1-integrin cell surface expression on the HR-3Y1-2 cells.
Decreased adhesion of HR-3Y1-2 cells to fibronectin-coated plates.
In Vivo

Apocynin (0.1 mg/kg/day, i.p., three times per week for 3 months) increases bone mineral density and total bone volume in SAMP6 mice[3].
Apocynin (5 mg/kg, i.p.) reduces the degree of lung injury and attenuates the degree of acute inflammation in the Carrageenan (HY-125474)-induced pleurisy mice[5].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: SAMP6 mouse model (Pharmacokinetic assay)[3]
Dosage: 0.1 mg/kg/day
Administration: Intraperitoneal injection (i.p.), three times per week, for 3 months
Result: Showed a higher bone value and exhibited a lower percentage of SA-β -gal positive cells than the control group.
Increased the expression of Ki67 and Oct-4 mRNA.
Altered the osteoblast-osteoclast balance in bone and promoted the activity of osteoblasts.
Animal Model: Carrageenan (HY-125474)-induced pleurisy in male adult CD1 mice [5]
Dosage: 5 mg/kg
Administration: Intraperitoneal injection (i.p.)
Result: Blocked NADPH oxidase activation and attenuated neutrophil infiltration and lipid peroxidation in the lung tissue.
Reduced the degree of PARP activation and the degree of IL-1b expression.
Prevented carrageenan-induced IkB-a degradation and reduced the levels of NF-kB p65.
Attenuated this iNOS expression,reduced the degree of positive staining for Fas ligand in the lung tissues.
Inhibited cells apoptosis in carrageenan-treated mice.
Prevented Bax expression and reduced the degree of positive staining for Bax.
Attenuated carrageenan-induced inhibition of Bcl-2 expression and the loss of positive staining for Bcl-2 in mice subjected to carrageenan-induced pleurisy.
Reduced the level of pERK1/2 and p38 expression.
Clinical Trial
Molecular Weight

166.17

Formula

C9H10O3
CAS No.
Appearance

Solid

Color

Off-white to gray

SMILES

CC(C1=CC=C(O)C(OC)=C1)=O
Structure Classification
Initial Source
Shipping

Room temperature in continental US; may vary elsewhere.
Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
In Vitro: 

DMSO : ≥ 100 mg/mL (601.79 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 3.33 mg/mL (20.04 mM; ultrasonic and warming and heat to 60°C)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 6.0179 mL 30.0897 mL 60.1793 mL
5 mM 1.2036 mL 6.0179 mL 12.0359 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (15.04 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (15.04 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Purity & Documentation
References

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