| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
BCIP targets alkaline phosphatase (ALP), an enzyme that catalyzes the hydrolysis of phosphate esters. As a chromogenic substrate, BCIP is specifically cleaved by alkaline phosphatase, releasing an indoxyl intermediate that undergoes oxidative dimerization to form an insoluble indigo dye. The compound does not have a therapeutic target but rather serves as a detection tool for alkaline phosphatase activity in biological samples. Its substrate specificity for alkaline phosphatase makes it a valuable reagent for enzyme activity assays and visualization of ALP-labeled probes in various detection systems.
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| ln Vitro |
BCIP exhibits no direct biological activity as a therapeutic agent; its primary function is as a chromogenic substrate for alkaline phosphatase. Upon enzymatic cleavage by alkaline phosphatase, BCIP produces a colored precipitate that can be visualized by light microscopy or measured spectrophotometrically. The compound's activity is entirely dependent on the presence of alkaline phosphatase, making it a sensitive and specific detection reagent for ALP-labeled antibodies, nucleic acid probes, or other ALP-conjugated molecules. In combination with NBT, the staining sensitivity is significantly amplified.
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| ln Vivo |
BCIP is not evaluated for in vivo activity in animal models as it is a laboratory reagent rather than a therapeutic agent. Its application is in diagnostic and research assays for the detection of alkaline phosphatase activity in tissue sections, cells, or biological fluids. The compound may be used ex vivo to stain tissue samples collected from animal studies for histopathological or immunohistochemical evaluation.
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| Enzyme Assay |
In vitro non-cell enzyme assays using BCIP involve incubating the substrate with alkaline phosphatase enzyme or ALP-conjugated probes in an appropriate buffer system. The enzymatic reaction releases an indoxyl intermediate that undergoes oxidative dimerization to form an insoluble indigo precipitate. The reaction can be monitored by colorimetric measurement of the precipitated dye or by absorbance measurement of the soluble product. For quantitative assays, the reaction is typically stopped and the absorbance is measured at 595 nm or other appropriate wavelengths. Standard curves are prepared using known amounts of alkaline phosphatase.
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| Cell Assay |
In vitro cell-based assays using BCIP are performed in immunohistochemistry (IHC), immunocytochemistry (ICC), or in situ hybridization (ISH) formats. Cells or tissue sections are incubated with alkaline phosphatase-conjugated antibodies or probes, followed by addition of BCIP substrate solution (often in combination with NBT). The enzymatic reaction produces a colored precipitate at the site of the target antigen or nucleic acid sequence. The staining is visualized by light microscopy, and the intensity of staining correlates with the abundance of the target molecule. Counterstaining is often performed to visualize cellular morphology.
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| Animal Protocol |
In vivo animal studies are not performed with BCIP as it is a diagnostic substrate rather than a therapeutic agent. The compound may be used ex vivo to stain tissue sections collected from animal studies for histopathological evaluation or to detect ALP activity in tissue samples. Standard immunohistochemistry or histochemistry protocols are followed, with tissue sections incubated with ALP-conjugated antibodies and BCIP/NBT substrate for color development.
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| ADME/Pharmacokinetics |
BCIP has a molecular weight of 433.62 g/mol and a molecular formula of C₈H₆BrClNO₄P·C₇H₉N. It has a melting point of 194-195°C, a boiling point of 580.2°C at 760 mmHg, and a LogP of 5.213. The compound is soluble in DMSO at ≥51 mg/mL (~117.61 mM) and in DMF at 25 mg/mL (~57.65 mM). It should be stored as a powder at -20°C for up to three years or at 4°C for up to two years, protected from moisture and light. In solvent, it can be stored at -80°C for six months or at -20°C for one month.
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| Toxicity/Toxicokinetics |
BCIP is considered a laboratory reagent with low toxicity at the concentrations used in staining procedures. However, standard laboratory safety precautions should be followed when handling the compound, including the use of appropriate personal protective equipment. The compound may cause skin and eye irritation. It is classified as a research reagent and is not intended for human therapeutic use. Comprehensive toxicological studies have not been performed as the compound is not used in vivo.
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| Additional Infomation |
BCIP is a chromogenic substrate for alkaline phosphatase, also known as 5-bromo-4-chloro-3-indolyl phosphate p-toluidine salt or X-phosphate p-toluidine salt. It is widely used in ELISA, immunohistochemistry, Western blotting, and in situ hybridization for the detection of alkaline phosphatase activity. The compound is often used in combination with NBT to amplify staining sensitivity. BCIP is available in two salt forms: the water-soluble disodium salt and the DMF-soluble p-toluidine salt. Not approved for therapeutic use; intended for research and diagnostic applications only.
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| Molecular Formula |
C8H6BRCLNO4P.C7H9N
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|---|---|
| Molecular Weight |
433.62
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| Exact Mass |
431.964
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| CAS # |
6578-06-9
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| PubChem CID |
81059
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| Appearance |
White to off-white solid powder
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| Boiling Point |
580.2ºC at 760 mmHg
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| Melting Point |
194-195ºC
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| Flash Point |
304.7ºC
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| LogP |
5.213
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
24
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| Complexity |
377
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
QEIFSLUFHRCVQL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C8H6BrClNO4P.C7H9N/c9-4-1-2-5-7(8(4)10)6(3-11-5)15-16(12,13)14;1-6-2-4-7(8)5-3-6/h1-3,11H,(H2,12,13,14);2-5H,8H2,1H3
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| Chemical Name |
(5-bromo-4-chloro-1H-indol-3-yl) dihydrogen phosphate;4-methylaniline
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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 : ≥ 51 mg/mL (~117.61 mM)
DMF : 25 mg/mL (~57.65 mM) |
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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 | 2.3062 mL | 11.5308 mL | 23.0617 mL | |
| 5 mM | 0.4612 mL | 2.3062 mL | 4.6123 mL | |
| 10 mM | 0.2306 mL | 1.1531 mL | 2.3062 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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