iE-DAP dihydrochloride

Cat. No.: HY-P5523A Purity: 99.72%: Data Sheet
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iE-DAP dihydrochloride is a Nod1 agonist. iE-DAP dihydrochloride activates NOD1, which in turn activates the NF-κB signaling pathway and MLCK signaling pathway, inducing cellular inflammatory responses and tight junction disruption. iE-DAP dihydrochloride downregulates the expression of ZO-1 and Occludin genes. iE-DAP dihydrochloride increases the secretion of IL-6, GRO-α, MCP-1, IL-8 and MIP-1β in term human trophoblast cell cultures. iE-DAP dihydrochloride triggers preterm birth in pregnant mice, reduces fetal body weight, and induces fetal inflammation. iE-DAP dihydrochloride can be used in studies related to mastitis and preterm birth.

For research use only. We do not sell to patients.

iE-DAP dihydrochloride Chemical Structure

Size	Stock	Quantity
1 mg	In-stock	Estimated Time of Arrival: December 31
5 mg	In-stock	Estimated Time of Arrival: December 31
10 mg	In-stock	Estimated Time of Arrival: December 31
50 mg	Get quote
100 mg	Get quote

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Other Forms of iE-DAP dihydrochloride:

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•Related Screening Libraries:

•Related Small Molecules:

•Related Proteins:

View All NOD-like Receptor (NLR) Isoform Specific Products:

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NLRP3 NLRP1 NOD1 NOD2

View All NF-κB Isoform Specific Products:

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NF-κB NF-κB1/p50 RelA/p65 RelB c-Rel

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IL-1 IL-2 IL-3 IL-4 IL-5 IL-6 IL-8 IL-10 IL-12 IL-13 IL-15 IL-17 IL-23 IL-7 IL-33 IL-22 IL-18

Biological Activity
Purity & Documentation
References
Customer Review

Description

In Vitro

iE-DAP dihydrochloride (10-10000 ng/mL; 1-24 h) upregulates the mRNA expression of IL-1β, IL-6 and IL-8 in BMECs, downregulates the mRNA expression of ZO-1 and Occludin in BMECs, reduces TEER, and increases the paracellular dextran permeability in BMECs^[1].
iE-DAP dihydrochloride (1000 ng/mL; 12 h) upregulates the protein expression levels of p-MLC2 and MLCK as well as the phosphorylation level of MLC2, downregulates the mRNA and protein expression levels of ZO-1 and Occludin, reduces TEER, increases the dextran permeability via the paracellular pathway, and disrupts the distribution of ZO-1 in BMECs^[1].
iE-DAP dihydrochloride (1000 ng/mL; 12 h) upregulates the mRNA expression and secretion levels of IL-1β, IL-6 and IL-8 in BMECs, and activates the NF-κB pathway by increasing the expression of p-p65 protein, the phosphorylation level of p65 and the nuclear translocation of p65^[1].
iE-DAP dihydrochloride (1000 ng/mL; 12 h) upregulates the expressions of NOD1 protein and p-p65 protein, enhances p65 phosphorylation level, and induces p65 nuclear translocation to activate the NOD1/NF-κB pathway^[1].
iE-DAP dihydrochloride (100 μg/mL; 72 h) potently stimulates the secretion of IL-6, GRO-α, MCP-1, IL-8 and MIP-1β by human primary third-trimester cytotrophoblast cells^[2].

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

Real Time qPCR^[1]

Cell Line:	bovine mammary epithelial cells (BMECs)
Concentration:	1000 ng/mL
Incubation Time:	1, 3, 6, 12, 24 h
Result:	Significantly upregulated IL-1β mRNA relative expression at 1 h. Significantly upregulated IL-6 mRNA relative expression at 1 h. Significantly upregulated IL-8 mRNA relative expression at 1 h, with all inflammatory cytokines increasing in a time-dependent manner. Significantly downregulated ZO-1 mRNA relative expression at 6 h. Significantly downregulated Occludin mRNA relative expression at 12 h, with both tight junction proteins decreasing in a time-dependent manner.

Real Time qPCR^[1]

Cell Line:	bovine mammary epithelial cells (BMECs)
Concentration:	0, 1, 10, 100, 1000, 10000 ng/mL
Incubation Time:	12 h
Result:	Increased the mRNA relative expression and release of IL-1β at 10 ng/mL. Increased the mRNA relative expression and release of IL-6 and IL-8 at 100 ng/mL. Increased the gene expression and release of inflammatory cytokines in a concentration-dependent manne. Reduced the relative mRNA expression of ZO-1、Occludin at 100、1000 ng/mL. Decreased the gene expression of TJ proteins in a concentration-dependent manne. Decreased the TEER and increased paracellular dextran passage at 100 ng/mL.

Western Blot Analysis^[1]

Cell Line:	bovine mammary epithelial cells (BMECs)
Concentration:	1000 ng/mL
Incubation Time:	12 h
Result:	Increased p-MLC2 protein expression and MLC2 phosphorylation, reduced protein expression of ZO-1 and Occludin, decreases TEER. Significantly increased p-p65 protein expression and the p-p65/p65 phosphorylation ratio. Significantly increased NOD1 protein expression. Significantly increased MLCK protein expression.

Immunofluorescence^[1]

Cell Line:	bovine mammary epithelial cells (BMECs)
Concentration:	1000 ng/mL
Incubation Time:	12 h
Result:	Caused redistribution of ZO-1 protein.\nInduced nuclear translocation of NF-κB p65.

In Vivo

iE-DAP dihydrochloride (500-1000 μg; i.p.; single dose) administered to pregnant C57BL/6J mice on embryonic day 14.5 induces 100% preterm delivery within 24 hours at 1000 μg, while the 750 μg dose reduces fetal weight and triggers maternal-fetal interface and fetal inflammation without inducing preterm delivery, and the 500 μg dose does not induce preterm delivery^[2].

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

Animal Model:	C57BL/6J mice (adult, 8-12 wk old, pregnant, preterm labor model induced at embryonic day 14.5)^[2]
Dosage:	500 μg; 750 μg; 1000 μg
Administration:	i.p.; single dose
Result:	Induced preterm delivery within 24 hours in 100% of treated mice (1000 μg dose). Reduced fetal weight, significantly increased placental Eotaxin levels, significantly decreased placental MCP-1 levels, significantly increased decidual IL-1α and RANTES levels, and triggered a significant fetal proinflammatory response with elevated levels of Eotaxin, IL-1β, IL-3, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-17, G-CSF, GM-CSF, MIP-1α, MIP-1β, KC, TNF-α, and IFN-γ (750 μg dose). Did not induce preterm delivery (500 μg dose).

Molecular Weight

392.23

Formula

C₁₂H₂₃Cl₂N₃O₇

Appearance

Solid

Color

White to off-white

Sequence

γ-d-Glu-meso-DAP

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Sealed storage, away from moisture

Powder	-80°C	2 years
	-20°C	1 year

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

Solvent & Solubility

In Vitro:

DMSO : 100 mg/mL (254.95 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	2.5495 mL	12.7476 mL	25.4952 mL
5 mM	0.5099 mL	2.5495 mL	5.0990 mL
10 mM	0.2550 mL	1.2748 mL	2.5495 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, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Molarity Calculator
Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

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 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (6.37 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.
Protocol 2

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (6.37 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 99.72%

Select Batch:

Data Sheet (279 KB) SDS (252 KB)

COA (246 KB) RP-HPLC (112 KB) MS (213 KB) Elemental Analysis Report (106 KB)

Handling Instructions (2659 KB)

References

[1]. Wang Y, et al. iE-DAP Induced Inflammatory Response and Tight Junction Disruption in Bovine Mammary Epithelial Cells via NOD1-Dependent NF-κB and MLCK Signaling Pathway. Int J Mol Sci. 2023;24(7):6263. Published 2023 Mar 27. [Content Brief]

[2]. Cardenas I, et al. Nod1 activation by bacterial iE-DAP induces maternal-fetal inflammation and preterm labor. J Immunol. 2011;187(2):980-986. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	2.5495 mL	12.7476 mL	25.4952 mL	63.7381 mL
	5 mM	0.5099 mL	2.5495 mL	5.0990 mL	12.7476 mL
	10 mM	0.2550 mL	1.2748 mL	2.5495 mL	6.3738 mL
	15 mM	0.1700 mL	0.8498 mL	1.6997 mL	4.2492 mL
	20 mM	0.1275 mL	0.6374 mL	1.2748 mL	3.1869 mL
	25 mM	0.1020 mL	0.5099 mL	1.0198 mL	2.5495 mL
	30 mM	0.0850 mL	0.4249 mL	0.8498 mL	2.1246 mL
	40 mM	0.0637 mL	0.3187 mL	0.6374 mL	1.5935 mL
	50 mM	0.0510 mL	0.2550 mL	0.5099 mL	1.2748 mL
	60 mM	0.0425 mL	0.2125 mL	0.4249 mL	1.0623 mL
	80 mM	0.0319 mL	0.1593 mL	0.3187 mL	0.7967 mL
	100 mM	0.0255 mL	0.1275 mL	0.2550 mL	0.6374 mL