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> X-GAL [7240-90-6]

X-GAL [7240-90-6]

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Cat# HY-15934-1g

Size : 1g

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X-GAL (BCIG) is a widely used chromogenic β-galactosidase substrate. X-GAL is a colorless compound until cleaved by β-galactosidase, at which point X-GAL turns to an insoluble and detectable blue compound, making X-GAL particularly useful in techniques such as blue-white screening for cloning in bacteria. X-GAL can also be used for detection of β-galactosidase activity.

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

X-GAL Chemical Structure

X-GAL Chemical Structure

CAS No. : 7240-90-6

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

Based on 1 publication(s) in Google Scholar

Other Forms of X-GAL:

  • 5-Bromo-4-chloro-3-indolyl-α-D-mannopyranoside, 93% Get quote
Description

X-GAL (BCIG) is a widely used chromogenic β-galactosidase substrate. X-GAL is a colorless compound until cleaved by β-galactosidase, at which point X-GAL turns to an insoluble and detectable blue compound, making X-GAL particularly useful in techniques such as blue-white screening for cloning in bacteria. X-GAL can also be used for detection of β-galactosidase activity[1][2].

In Vitro

When the cells (HUVEC for example) were grown in dishes to 80% confluence, 50 μmol/L H2O2 were added in the medium for 12 h.
Subsequently, cells were harvested with trypsin/EDTA and washed three times with PBS and further fixed with 4% formaldehyde for 10 min.
As control, cells without H2O2 treatment were prepared in the same way. Cells were stained with by X-gal at 37 ℃[2].

The intestinal tissue was excised and cut longitudinally. After the intestinal content was rinsed off with phosphate buffer solution, the intestinal tissue was placed with the inner surface facing up on the slide.
Next, a single blue colony containing β-gal (E. coli DH5α containing pUC18) was dilution in 100 μL PBS and then 10 μL were added in the tissue.
After incubation for 1 h, the tissues were washed with PBS solution.
As control, tissues without the blue colony were prepared in the same way.
All tissues were fixed with 4% formaldehyde for 10 min and were further stained by adding X-gal at 37 ℃[2].

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

X-GAL Related Antibodies
Molecular Weight

408.63

Formula

C14H15BrClNO6
CAS No.

7240-90-6
Appearance

Solid

Color

White to off-white

SMILES

O[C@H]([C@@H](O)[C@H]1O)[C@@H](O[C@@H]1CO)OC2=CNC3=C2C(Cl)=C(Br)C=C3
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMF : 200 mg/mL (489.44 mM; Need ultrasonic)

DMSO : 100 mg/mL (244.72 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 2.4472 mL 12.2360 mL 24.4720 mL
5 mM 0.4894 mL 2.4472 mL 4.8944 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 (protect from 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 (5.09 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.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (5.09 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 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.
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 (protect from 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

Purity: 99.88%

COA (249 KB) HNMR (188 KB) LCMS (484 KB)

Handling Instructions (2659 KB)
References

  • [1]. Sanchez-Ramos J, et al. The X-gal caution in neural transplantation studies. Cell Transplant. 2000 Sep-Oct;9(5):657-67.  [Content Brief]

    [2]. Li S, et al. In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times. Talanta. 2021 Nov 1;234:122689.  [Content Brief]

  • [1]. Sanchez-Ramos J, et al. The X-gal caution in neural transplantation studies. Cell Transplant. 2000 Sep-Oct;9(5):657-67.

    [2]. Li S, et al. In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times. Talanta. 2021 Nov 1;234:122689.

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 (protect from 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 / DMF 1 mM 2.4472 mL 12.2360 mL 24.4720 mL 61.1800 mL
5 mM 0.4894 mL 2.4472 mL 4.8944 mL 12.2360 mL
10 mM 0.2447 mL 1.2236 mL 2.4472 mL 6.1180 mL
15 mM 0.1631 mL 0.8157 mL 1.6315 mL 4.0787 mL
20 mM 0.1224 mL 0.6118 mL 1.2236 mL 3.0590 mL
25 mM 0.0979 mL 0.4894 mL 0.9789 mL 2.4472 mL
30 mM 0.0816 mL 0.4079 mL 0.8157 mL 2.0393 mL
40 mM 0.0612 mL 0.3059 mL 0.6118 mL 1.5295 mL
50 mM 0.0489 mL 0.2447 mL 0.4894 mL 1.2236 mL
60 mM 0.0408 mL 0.2039 mL 0.4079 mL 1.0197 mL
80 mM 0.0306 mL 0.1530 mL 0.3059 mL 0.7648 mL
100 mM 0.0245 mL 0.1224 mL 0.2447 mL 0.6118 mL
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X-GAL Related Classifications

  • Others
  • Others Metabolic Enzyme/Protease
  • Fluorescent Dye Glucosidase
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:

X-GAL7240-90-6BCIGFluorescent DyeGlucosidaseβ-galactosidase substrateblue-white screeningInhibitorinhibitorinhibit

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