| Size | Price | Stock | Qty |
|---|---|---|---|
| 250mg |
|
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| Other Sizes |
| Targets |
Alkaline phosphatase (ALP) is the primary molecular target. The phosphate group on BCIP is specifically hydrolyzed by ALP, releasing the indolyl intermediate which then oxidizes to form a colored product. No other phosphatases typically produce this reaction.
|
|---|---|
| ln Vitro |
For life science-related study, sodium 5-bromo-4-chloro-1H-indol-3-yl phosphate is a biochemical reagent that can be utilized as an organic substance or biological material.
In cell-free assays, BCIP is highly sensitive and specific for ALP. At 37degC in Tris buffer (pH 9.0-9.5), BCIP is rapidly dephosphorylated by ALP (activity as low as 0.01 U/mL can be detected). The reaction produces a deep blue precipitate (λmax 615 nm) which can be measured spectrophotometrically or visually. |
| ln Vivo |
There is no in vivo activity for BCIP as it is not used in live animals. It is a reagent for in vitro histochemical and blotting applications. No systemic activity is reported.
|
| Enzyme Assay |
Nitrocellulose membranes with immobilized ALP-conjugated antibodies are incubated with BCIP (0.15 mg/mL) and NBT (0.3 mg/mL) in alkaline phosphatase buffer (100 mM Tris-HCl pH 9.5, 100 mM NaCl, 5 mM MgCl2) for 5-30 minutes at room temperature in the dark. The reaction is stopped by washing with water, producing purple bands.
|
| Cell Assay |
For cellular staining, cells or tissue sections fixed on slides are incubated with ALP-conjugated primary or secondary antibody, washed, then incubated with BCIP/NBT substrate solution (prepared as above) for 10-60 minutes at 37degC. Positive staining appears as blue-purple precipitate. Counterstaining can be performed with nuclear fast red.
|
| Animal Protocol |
There is no in vivo animal protocol for BCIP. It is used ex vivo on tissue sections. For whole-mount staining, embryos or tissues are fixed, permeabilized, incubated with ALP-conjugated probes, then stained with BCIP/NBT solution for several hours to overnight, followed by clearing and imaging.
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| ADME/Pharmacokinetics |
PK properties are not applicable as BCIP is not administered systemically. It is a labile substrate; stock solutions (10 mg/mL in DMF) should be stored at -20degC and protected from light. Working solutions in alkaline buffer are stable for a few hours.
|
| Toxicity/Toxicokinetics |
General toxicity: BCIP is of low acute toxicity. The product is an irritant to eyes and skin. Safety data sheets recommend avoiding inhalation or ingestion. Standard lab safety precautions (gloves, goggles) should be used. It is not for human consumption.
|
| Additional Infomation |
See other relationships...
BCIP is commonly used in conjunction with NBT (nitro blue tetrazolium) in a BCIP/NBT system for Western blot, Southern blot, immunohistochemistry, and in situ hybridization. The reaction produces a stable, insoluble purple precipitate that does not fade. This product is for research use only. |
| Molecular Formula |
C8H4BRCLNNA2O4P
|
|---|---|
| Molecular Weight |
370.43
|
| Exact Mass |
368.854
|
| CAS # |
102185-33-1
|
| PubChem CID |
6097197
|
| Appearance |
White to off-white solid powder
|
| Boiling Point |
580.2ºC at 760 mmHg
|
| Melting Point |
300°C
|
| Flash Point |
304.7ºC
|
| Vapour Pressure |
2.65E-14mmHg at 25°C
|
| LogP |
3.931
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
18
|
| Complexity |
303
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC(=C(C2=C1NC=C2OP(=O)([O-])[O-])Cl)Br.[Na+].[Na+]
|
| InChi Key |
OAZUOCJOEUNDEK-UHFFFAOYSA-L
|
| InChi Code |
InChI=1S/C8H6BrClNO4P.2Na/c9-4-1-2-5-7(8(4)10)6(3-11-5)15-16(12,13)14;;/h1-3,11H,(H2,12,13,14);;/q;2*+1/p-2
|
| Chemical Name |
disodium;(5-bromo-4-chloro-1H-indol-3-yl) phosphate
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.6996 mL | 13.4978 mL | 26.9957 mL | |
| 5 mM | 0.5399 mL | 2.6996 mL | 5.3991 mL | |
| 10 mM | 0.2700 mL | 1.3498 mL | 2.6996 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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