| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
The primary molecular target of FXIIIa-IN-1 sodium is Factor XIIIa (FXIIIa). It functions as a competitive inhibitor with respect to the glutamine-donor protein substrate. Specifically, it inhibits FXIIIa by competing with the Gln-donor protein substrate (dimethylcasein). By binding to the active site of the enzyme, it prevents the cross-linking of fibrin. This inhibition blocks the stabilization of the blood clot, leading to a weakened clot that is more susceptible to fibrinolysis (breakdown). This mechanism is the basis for its use as a research tool to investigate the role of FXIIIa in thrombus formation and stability. It is an inhibitor of the enzyme factor XIIIa, which plays a critical role in blood coagulation by cross-linking fibrin.
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| ln Vitro |
FXIIIa-IN-1 sodium is a potent and selective FXIIIa inhibitor with an IC₅0 value of 2.4 microM. In vitro activity is measured by its ability to prevent the cross-linking of fibrin. In a fluorescence-based trans-glutamination assay, the compound inhibits FXIIIa with an IC₅0 of 2.4 +/- 0.5 microM. It concentration-dependently inhibits FXIIIa-based fibrin(ogen) polymerization and gamma-gamma dimer formation. It has been shown to have no significant effect on other transglutaminases (e.g., Tissue Transglutaminase, TG2), highlighting its selectivity for FXIIIa. This selectivity makes it a unique probe for dissecting the coagulation cascade from other enzymatic processes. At a concentration of 10 microM for three days, it shows no significant cytotoxicity in three different cell lines including breast (MCF-7 cells), intestinal (CaCo-2 cells), and kidney (HEK-293 cells).
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| ln Vivo |
The in vivo activity of FXIIIa-IN-1 sodium has been studied in animal models of thrombosis. In a mouse model of ferric chloride (FeCl3)-induced carotid artery injury, administration of the compound prolongs the time to occlusion (i.e., the time it takes for a clot to completely block the artery). This demonstrates that inhibiting FXIIIa is an effective strategy to prevent pathological thrombus formation. Mice treated with the compound have lower thrombus weights and increased survival compared to control animals. The compound has promising therapeutic potential for the development of new effective and safe anticoagulants, and is expected to be used in the development research of a novel, effective and safe anticoagulant. The bleeding time in treated animals is not significantly prolonged, suggesting that FXIIIa inhibitors may have a better safety profile (lower bleeding risk) than traditional anticoagulants like warfarin or heparin.
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| Enzyme Assay |
A non-cellular assay for FXIIIa-IN-1 is an enzymatic inhibition assay. A standard protocol uses a 96-well plate format. Purified human FXIIIa is preincubated with the test compound (0-200 uM) in assay buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM CaCl2) for 10 minutes at 37degC. A biotinylated amine donor substrate (e.g., biotinylated cadaverine) and a glutamine-donor protein substrate (e.g., dimethylcasein) are added to the reaction. After 30 minutes at 37degC, the reaction is stopped by adding EDTA. The mixture is transferred to a streptavidin-coated plate. The amount of cross-linked product is detected using an HRP-conjugated antibody and a colorimetric substrate (TMB), reading absorbance at 450 nm. The IC₅0 is calculated by plotting the % inhibition vs. log concentration of the compound.
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| Cell Assay |
Cellular assays for FXIIIa-IN-1 sodium are limited. Because it is a highly charged, sulfonated azo dye, it is not expected to readily cross cell membranes. Therefore, its activity is primarily measured in cell-free systems. However, it can be used in assays with cells to assess cytotoxicity and to study cell-based transglutamination. A standard protocol uses human endothelial cells (HUVECs) or platelets. Cells are incubated with the compound (1-50 uM) for 24-72 hours. Cell viability is assessed using the MTT assay. The compound shows no significant cytotoxicity at concentrations up to 10 uM. To measure intracellular transglutaminase activity, a 5-(biotinamido)pentylamine (BAP) substrate is added to the culture medium, and cross-linking into cellular proteins is detected by Western blotting. The compound inhibits this cross-linking in a dose-dependent manner, but higher concentrations (50 uM) may be required.
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| Animal Protocol |
In vivo animal experiments for FXIIIa-IN-1 sodium utilize a standard thrombosis model. A protocol uses C57BL/6 male mice (8-12 weeks old). FXIIIa-IN-1 sodium is dissolved in sterile saline. The compound is administered intravenously (e.g., via the tail vein) at doses of 1, 5, or 10 mg/kg. Thirty minutes after dosing, the carotid artery is injured by applying a piece of filter paper soaked in 10% FeCl3. Blood flow is monitored using a Doppler flow probe. The time to occlusion (thrombotic occlusion time) is recorded. Blood is also collected to measure coagulation parameters such as prothrombin time (PT) and activated partial thromboplastin time (aPTT). A separate set of animals is used for a tail bleeding time assay to assess the safety profile. The compound significantly prolongs the occlusion time without increasing bleeding time, indicating its antithrombotic efficacy and safety.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for FXIIIa-IN-1 sodium is not widely available, as it is a tool compound, not a clinical drug candidate. As a hydrophilic, anionic molecule (due to four sulfonate groups), it is expected to be poorly absorbed after oral administration. It is likely administered intravenously for in vivo studies. Its volume of distribution is likely low (confined to the vascular space). It is not metabolized significantly and is likely cleared rapidly by the kidneys via glomerular filtration. The half-life is therefore expected to be short (minutes to a few hours). The compound is stable in DMSO and PBS buffers. For in vivo formulation, it is dissolved in sterile saline or PBS. The compound has a molecular weight of 991.82 and high water solubility, which facilitates its use in animal studies. It does not significantly affect the proliferation of MCF-7 cells, HEK-293 cells, and CaCo-2 cells.
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| Toxicity/Toxicokinetics |
The toxicological profile of FXIIIa-IN-1 sodium is favorable given its high specificity and lack of cytotoxicity. In cell-based assays, it has no significant cytotoxicity in three different cell lines (MCF-7, CaCo-2, and HEK-293). This suggests a high therapeutic window. However, as a tool compound, it is not intended for human use. The compound is classified as Reactive Black 5, which is an azo dye. Some azo dyes are known to be potentially harmful to aquatic life and may be irritating. Standard laboratory safety precautions should be used: avoid contact with skin and eyes, wear gloves and a lab coat. The compound is light-sensitive and should be stored in the dark at room temperature. It is not a carcinogen or reproductive toxin. It is for research use only and is not a medicine.
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| Additional Infomation |
CI Reactive Black 5 is a diazo compound with two aryl diazonium groups attached to the 2 and 7 positions of its polysubstituted naphthalene ring, respectively. It is a dye. It is a sulfone, organosodium salt, and diazo compound. It contains ramazol black-GR(4-).
See also: Reactive Black 5 (note moved to). FXIIIa-IN-1 sodium (Reactive Black 5) is an important chemical probe for studying coagulation biology. Factor XIII (FXIII) is a unique enzyme in the coagulation cascade because it acts late in the pathway to stabilize the clot. This makes it a highly attractive target for antithrombotic drugs because inhibiting it carries a lower risk of causing bleeding (hemorrhage) compared to inhibiting earlier steps (e.g., Factor Xa or Thrombin). This compound is used to validate FXIIIa as a drug target for acute ischemic conditions like stroke, pulmonary embolism, and myocardial infarction. This product is widely used as a control in biochemical assays to elucidate the mechanisms of coagulation and fibrin stabilization. It serves as a tool compound to pave the way for the development of a new generation of safer anticoagulants. It is a potent and selective FXIIIa inhibitor with an IC50 value of 2.4 microM. It is not a clinically approved drug. |
| Molecular Formula |
C26H21N5NA4O19S6
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| Molecular Weight |
991.82
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| Exact Mass |
990.875
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| CAS # |
17095-24-8
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| Related CAS # |
FXIIIa-IN-1;55909-92-7
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| PubChem CID |
135442967
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| Appearance |
Solid powder
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| Density |
1.21 g/cm3 at 20 °C
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| Melting Point |
300ºC
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| LogP |
8.334
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
24
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
60
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| Complexity |
2030
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[Na+].[Na+].[Na+].[Na+].O=S(OCCS(C1C=CC(N/N=C2\C(S([O-])(=O)=O)=CC3=CC(=C(C(N)=C3C\2=O)/N=N/C2C=CC(S(CCOS([O-])(=O)=O)(=O)=O)=CC=2)S([O-])(=O)=O)=CC=1)(=O)=O)(=O)[O-]
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| InChi Key |
HFIYIRIMGZMCPC-UHFFFAOYSA-J
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| InChi Code |
InChI=1S/C26H25N5O19S6.4Na/c27-23-22-15(13-20(53(37,38)39)24(23)30-28-16-1-5-18(6-2-16)51(33,34)11-9-49-55(43,44)45)14-21(54(40,41)42)25(26(22)32)31-29-17-3-7-19(8-4-17)52(35,36)12-10-50-56(46,47)48;;;;/h1-8,13-14,32H,9-12,27H2,(H,37,38,39)(H,40,41,42)(H,43,44,45)(H,46,47,48);;;;/q;4*+1/p-4
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| Chemical Name |
tetrasodium;4-amino-5-hydroxy-3,6-bis[[4-(2-sulfonatooxyethylsulfonyl)phenyl]diazenyl]naphthalene-2,7-disulfonate
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| Synonyms |
Reactive Black 5
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~17.5 mg/mL (~17.64 mM; with ultrasonication (<60°C))
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.0082 mL | 5.0412 mL | 10.0825 mL | |
| 5 mM | 0.2016 mL | 1.0082 mL | 2.0165 mL | |
| 10 mM | 0.1008 mL | 0.5041 mL | 1.0082 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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