2. Others Metabolic Enzyme/Protease
  3. Fluorescent Dye Glycosidase
  4. X-GAL

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.

CAS No. : 7240-90-6

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Customer Review

Based on 5 publication(s) in Google Scholar

Other Forms of X-GAL:

X-GAL Related Antibodies

Top Publications Citing Use of Products

    X-GAL purchased from MedChemExpress. Usage Cited in: Cell. 2025 Sep 18;188(19):5403-5425.e33.  [Abstract]

    X-gal (1 mg/mL; 37 ℃; 12–72 h). SA-β-Gal staining of hMACs treated with Veh, IgG, Bet, or IgG + Bet. The percentage of positive cells is quantified.

    X-GAL purchased from MedChemExpress. Usage Cited in: Johannes Kepler University Linz. 2025.

    X-GAL (37℃; 3.5 h). Senescence-associated-β-Galactosidase staining images of untreated and treated K52-pBabe-empty BSD and K562-pBabe-3xFlag-BSD-METTL3 cells.

    X-GAL purchased from MedChemExpress. Usage Cited in: Nat Aging. 2023 Feb;3(2):185-201.

    X-GAL (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) (37 ℃; 1 mg/mL; overnight). Representative images of SA-β-gal positive cells in IMR90 cells were shown. SA-β-gal positive cells (%) reduced.

    X-GAL purchased from MedChemExpress. Usage Cited in: Nat Aging. 2023 Feb;3(2):185-201.

    X-GAL (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) (37 ℃; 1 mg/mL; overnight). β-Gal staining analysis of HGADFN003 cells with the indicated treatments. Representative images and quantification of SA-β-gal positive cells (%).

    X-GAL purchased from MedChemExpress. Usage Cited in: University of Jena. 2023 Dec.

    X-gal (1 mg/mL; 37 ℃; 8 h). Senescence-associated β-galactosidase staining (SABG) was performed.

    • Biological Activity

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    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

    Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
    1. Solution preparation[1]
    1.1 Preparation of stock solution
    Solvent: DMF or DMSO.
    Concentration: 20 mg/mL is recommended.
    Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
    1.2 Preparation of working solution
    Dilute X-gal to 40 μg/mL with 1 mL agar medium, and add 1 μL of IPTG (HY-15921) (24 mg/mL) and 1 μL of AMP (HY-A0181) (100 mg/mL) (optimized according to the experiment).
    Note: The working solution should be prepared and used immediately. Keep it away from light.
    2. Blue/white colony screening
    2.1 After the DNA fragment is inserted into the pUC series vector (or other vectors with lacZ, Amp genes) and then transformed into lacZ-deficient cells, apply the above-mentioned X-gal, IPTG, and AMP plate culture medium.
    2.2 Incubate at 37°C until blue-white colonies appear on the agar surface.
    2.3 Cultivate the blue-white colonies further and identify them by double enzyme digestion.
    2.4 Aspirate the dye working solution, wash with PBS 2-3 times, and observe under a microscope.
    3. Cellular level detection[2]
    3.1 When cells are cultured to 80% confluence in culture dishes, add 50 μmol/L H2O2 to the medium and incubate for 12 h.
    3.2 Harvest cells with trypsin/EDTA, wash three times with PBS, and fix with 4% formaldehyde for 10 min.
    3.3 Prepare cells without H2O2 treatment in the same way as the control group.
    3.4 Perform X-gal staining at 37 ℃.
    4. Tissue-level detection[2]
    4.1 Excise intestinal tissues and cut them longitudinally. Rinse off intestinal contents with phosphate buffered saline, then place the intestinal tissues on glass slides with the inner surface facing upward.
    4.2 Dilute a single blue colony containing β-gal (Escherichia coli DH5α carrying pUC18) in 100 μL PBS, and then add 10 μL of the dilution to the tissues.
    4.3 After incubating for 1 hour, wash the tissues with PBS solution. For the control group, prepare tissues without blue colonies using the same method.
    4.4 Fix all tissues with 4% formaldehyde for 10 minutes, and further stain with X-gal at 37 ℃.

    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.
    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)

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

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    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

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    (per animal)

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    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Calculation results:
    Working solution concentration: mg/mL
    Purity & Documentation

    Purity: 99.88%

    SDS (393 KB)

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

    Handling Instructions (2659 KB)
    References

    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

    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 DyeGlycosidaseβ-galactosidase substrateblue-white screeningInhibitorinhibitorinhibit

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