Alcian Blue 8GX [33864-99-2]

Cat# HY-D0001-50mg

Size : 50mg

Brand : MedChemExpress


Alcian Blue 8GX is a commonly used phthalocyanine dye that binds to glycoproteins and glycosaminoglycans. Alcian Blue 8GX has a wide range of applications in biological staining, including proteins in brain tumors and DNA in cells and tissues.

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

Alcian Blue 8GX Chemical Structure

Alcian Blue 8GX Chemical Structure

CAS No. : 33864-99-2

This product is a controlled substance and not for sale in your territory.

Based on 3 publication(s) in Google Scholar

Description

Alcian Blue 8GX is a commonly used phthalocyanine dye that binds to glycoproteins and glycosaminoglycans. Alcian Blue 8GX has a wide range of applications in biological staining, including proteins in brain tumors and DNA in cells and tissues[1][2][3][4].

In Vitro

Alcian Blue 8GX working solution preparation[1]
To prepare 1 mg/mL Alcian Blue 8GX: add 0.6 mL of glacial acetic acid to ddH2O to a final volume of 20 mL. Add 20 mg of Alcian Blue 8GX, dissolve thoroughly.
Alcian Blue 8GX staining of cell micromass cultures[1]
(1) Prepare preprocessing solution A: add 0.5 mL of ammonium hydroxide solution to 500 mL of 70% ethanol.
(2) Prepare preprocessing solution B: add 25 mL of glacial acetic acid to 70% ethanol to a final volume of 500 mL.
(3) Discard the culture medium from the micromass culture and fix with 1 mL of Bouin's fixative at room temperature for 15 minutes.
(4) Wash with preprocessing solution A and B.
(5) Stain with 2 mL of 1 mg/mL Alcian Blue 8GX at room temperature for 1 hour.
(6) Wash off the dye with deionized water, repeat four times. and soak in 2 mL of deionized water.
(7) Take a photograph.
(8) Wash off the dye again with deionized water.
(9) Extract the dye by incubating the culture with 1 mL of 6 M guanidine hydrochloride at room temperature overnight.
(10) Measure the optical density at OD595 using a spectrophotometer.
378/2000 Cell Culture Precautions[1]
(1) Avoiding Cell Death: To prevent cell death, embryos/cells should always be kept on ice before plating.
(2) Handling When Cell Viability is Below 70-80%: If cell viability drops below 70-80%, consider using a dead cell removal kit (e.g., Miltenyi Dead Cell Removal Kit, Miltenyi Biotech, Cat. No: 130-090-101) to eliminate dead cells.
(3) Cell Plating in 35 mm Culture Dishes: When plating cells in 35 mm culture dishes, maintain a high-density cell area. After 1.5 hours of culture, the cells should adhere. Carefully add high DMEM culture medium containing fibronectin.
(4) Use and Passaging of Isolated Cells: Isolated cells can be used for micro-clump culture on the same day they are isolated. However, these cells can only be passaged a maximum of three times, and the total culture time from isolation should not exceed 10 days.
Alcian Blue 8GX staining of glycosaminoglycans[2]
(1) Mast cells were fixed with methanol-40% formaldehyde-acetic acid (85:10:5 volume ratio) for 30 min, and the cells were stained after washing.
(2) Stain with 0.5% (w/v) Alcian Blue 8GX dissolved in 3% (v/v) acetic acid, pH 2.4, for 0.5 or 2 hours.
(3) The same solution saturated with urea as in (1) was returned to pH 2.4 with HCl and stained for 0.5 h.
(4) After staining with the same solution as in (1) for 0.5 h, the slides were stained with 0.3% (w/v) hematoxylin O (pH 0.9) in 0.125 N HCl for 5 min.
(5) After staining, rinse with distilled water, let it air dry, and then mount with Entellan.
(6) To detect the presence of histamine, air dry the unfixed or fixed smears of rat peritoneal mast cells, and stain them with 1% (w/v) ortho-phthalaldehyde (OPT) dissolved in xylene (staining time 2 minutes). The staining process is carried out in a closed chamber at room temperature with 100% relative humidity. After staining, the slides are drained and immediately mounted with tetrahydrofuran.
(7) The yellow fluorescence intensity of the histamine-OPT complex in mast cell granules was examined using a Leitz Orthoplan microscope equipped with a Leitz Ploemopak 2 (for incident fluorescence) and a 75 W high-pressure xenon lamp. The filter system used consisted of a TK 510/K 515 dichroic, a K 510 blocking filter and a 2 x KP 490 (= KP 500) excitation filter.
Alcian Blue 8GX staining of fetal cartilage[3]
(1) Rat fetuses were obtained from mated female Spraque-Dawley rats. On gestational day 21 (sperm/sperm day 5, gestational day 0), rats were euthanized by carbon dioxide asphyxiation. Fetuses were obtained by standard cesarean section and subsequently euthanized by oral administration of sodium pentobarbital. New Zealand white and Dutch rabbits were 5-6 months old at the time of mating, and pregnant rabbits were sacrificed on gestational day 28 and fetuses were obtained by intravenous injection of sodium pentobarbital. Each fetus was euthanized by oral administration of sodium pentobarbital.
(2) Fetuses were eviscerated, skinned, and fixed in 70% ethanol for 3-7 days.
(3) Alcian Blue 8GX was added to a mixture of 8 parts 95% ethanol and 2 parts glacial acetic acid to prepare solutions of 75 and 100 mg/L.
(4) Rat fetuses were stained in Alcian Blue 8GX (100 mg/L) for 24 h and rabbit fetuses were stained in Alcian Blue 8GX (75 mg/L) for 24 h, followed by rinsing in 95% ethanol for 6 h.
(5) Rat and rabbit fetuses were immersed in 1% and 1.25% potassium hydroxide, respectively. Rat fetuses were stained with 12 mg/L Alizarin Red S (HY-120601) for 48 h and rabbit fetuses were stained with 6 mg/L Alizarin Red S (HY-120601) for 24 h.
(6) Transferred to a solution of 70% ethanol and glycerol (1:1) for clearing, hardening, and examination.
Alcian Blue 8GX for protein determination[4]
(1) Dissolve the target protein in water or 0.1 M NaCl solution to a working concentration of 25 μg/ml.
(2) Use Britton-Robinson buffer (pH 7.24, containing 0.04 M H3PO4, 0.04 M HAc and 0.04 M H3BO3) to control the acidity and use 0.5 M NaCl solution to adjust the ionic strength of the aqueous solution.
(3) Add the appropriate protein working solution, 1.0 mL of Britton-Robinson buffer. Except for IgG, which was diluted with 0.1 M NaCl, all other mixtures were diluted to 10 mL with double distilled water.
(4) The mixture was used for absorption or RLS measurement. The RLS spectrum was scanned by adjusting the excitation and emission monochromators (∆λ = 0 nm) of the RF-540 fluorescence spectrometer. The RLS intensity was measured at 398.0 nm, proteins, including bovine serum albumin (BSA), human serum albumin (HAS), pepsin (Pep) and cellulase (Cel), can be determined with the limit of detection below 30 ng/mL. (5) RLS spectra of the interaction between ABGX and BSA. Concentration 1.0 × 10-5 M.

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

Molecular Weight

1298.86

Formula

C56H68Cl4CuN16S4

CAS No.

33864-99-2

Appearance

Solid

Color

Dark purple to black

SMILES

CN(C)/C(SCC(C=C1)=CC2=C1/C3=N/C(C4=C/5C=CC(CS/C(N(C)C)=[N+](C)\C)=C4)=NC5=N/C6=C(C=C(CS/C(N(C)C)=[N+](C)\C)C=C7)C7=C8N6[Cu]N3/C2=N\C9=N/C(C%10=C9C=CC(CS/C(N(C)C)=[N+](C)/C)=C%10)=N\8)=[N+](C)\C.[Cl-].[Cl-].[Cl-].[Cl-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

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

Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (76.99 mM; ultrasonic and warming and heat to 60°C; 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 0.7699 mL 3.8495 mL 7.6991 mL
5 mM 0.1540 mL 0.7699 mL 1.5398 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 and light). 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)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

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    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.08 mg/mL (1.60 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

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 and light)

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

References
  • [1]. Hiroki Ueharu, et al. Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice. Bio Protoc. 2022 Apr 5;12(7):e4371.  [Content Brief]

    [2]. J Tas, et al. The Alcian blue and combined Alcian blue--Safranin O staining of glycosaminoglycans studied in a model system and in mast cells. Histochem J. 1977 Mar;9(2):205-30.  [Content Brief]

    [3]. Brian G Redfern, et al. An alternative Alcian Blue dye variant for the evaluation of fetal cartilage. Birth Defects Res B Dev Reprod Toxicol. 2007 Jun;80(3):171-6.  [Content Brief]

    [4]. Yuan Fang Li, et al. Determination of Proteins Based on Their Resonance Light Scattering Enhancement Effect on Alcian Blue 8GX. Original Papers. 2000 Dec; VOL.16

  • [1]. Hiroki Ueharu, et al. Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice. Bio Protoc. 2022 Apr 5;12(7):e4371.

    [2]. J Tas, et al. The Alcian blue and combined Alcian blue--Safranin O staining of glycosaminoglycans studied in a model system and in mast cells. Histochem J. 1977 Mar;9(2):205-30.

    [3]. Brian G Redfern, et al. An alternative Alcian Blue dye variant for the evaluation of fetal cartilage. Birth Defects Res B Dev Reprod Toxicol. 2007 Jun;80(3):171-6.

    [4]. Yuan Fang Li, et al. Determination of Proteins Based on Their Resonance Light Scattering Enhancement Effect on Alcian Blue 8GX. Original Papers. 2000 Dec; VOL.16

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 and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 0.7699 mL 3.8495 mL 7.6991 mL 19.2476 mL
5 mM 0.1540 mL 0.7699 mL 1.5398 mL 3.8495 mL
10 mM 0.0770 mL 0.3850 mL 0.7699 mL 1.9248 mL
15 mM 0.0513 mL 0.2566 mL 0.5133 mL 1.2832 mL
20 mM 0.0385 mL 0.1925 mL 0.3850 mL 0.9624 mL
25 mM 0.0308 mL 0.1540 mL 0.3080 mL 0.7699 mL
30 mM 0.0257 mL 0.1283 mL 0.2566 mL 0.6416 mL
40 mM 0.0192 mL 0.0962 mL 0.1925 mL 0.4812 mL
50 mM 0.0154 mL 0.0770 mL 0.1540 mL 0.3850 mL
60 mM 0.0128 mL 0.0642 mL 0.1283 mL 0.3208 mL
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Alcian Blue 8GX Related Classifications

  • Others
  • Others
  • Fluorescent Dye
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Alcian Blue 8GX33864-99-2Fluorescent Dyecationic copper phthalocyanine dyecounterstaincell shapesInhibitorinhibitorinhibit