beta Galactosidase Antibody [FITC]

Western Blot: beta Galactosidase Antibody [FITC] [NB120-6641] - Lane 1 shows 80 ng and lane 2 shows 20 ng loaded onto gel. Results for non-reducing conditions of SDS-PAGE prior to transfer to nitrocellulose are shown on the left side of the figure; results obtained under reducing conditions are shown on the right. Blots were blocked overnight at 4 C with Blocking Buffer for Fluorescent Western Blotting. The membrane was probed with anti-b-Galactosidase diluted to 1:10,000. Reaction occurred overnight at 4C. Dylight649 conjugated Gt-a-anti-Rabbit IgG was used for detection.

Western Blot: beta Galactosidase Antibody [FITC] [NB120-6641] - Analysis of a band at 117 kDa (lane 1) corresponding to b-Gal present in a partially purified preparation (arrowhead). Approximately 1ug of protein was resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1,000. Reaction occurred overnight at 4 C followed by washes and reaction with a 1:10,000 dilution of IRDye 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 45 min at room temperature (800 nm channel, green).

Western Blot: beta Galactosidase Antibody [FITC] [NB120-6641] - Shows a band at 117 kDa (lanes 1 - 3) corresponding to 60 ng, 30 ng and 15 ng, respectively of b-Gal present in partially purified preparations (arrowhead). Lane 4 shows no cross reactivity with proteins present in a non-specific control E.coli lysate. Proteins were resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred to nitrocellulose and blocking using Blocking Buffer for Fluorescent Western Blotting. The membrane was probed with fluorescein conjugated anti-b-Galactosidase diluted to 1:10,000. Reaction occurred for 2 hours at room temperature.

Western blotting using Fluorescein conjugated anti-b-Galactosidase antibody shows a band at ~117 kDa (lanes 1 - 3) corresponding to 60 ng, 30 ng and 15 ng, respectively of b-Gal present in partially purified preparations (arrowhead). Lane 4 shows no cross reactivity with proteins present in a non-specific control E.coli lysate. Proteins were resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred to nitrocellulose and blocking using Blocking Buffer for Fluorescent Western Blotting

• Reactivity: Mu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: FITC

beta Galactosidase Antibody [FITC] Summary

• Description: For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room
  
  This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-fluorescein, anti-Rabbit Serum and purified and partially purified Beta Galactosidase [E.coli]
• Immunogen: Full length native beta Galactosidase isolated from E.coli
• Localization: Cytoplasmic
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:500-1:2500
  • Western Blot 1:10000
• Application Notes: This product has been tested by western blot and is designed for fluorescent western blotting, also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.

Reactivity Notes

Cross reactivity against beta Galactosidase from other tissues and species may occur but have not been specifically determined

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Alternate Names for beta Galactosidase Antibody [FITC]

• EC 3.2.1.23; b-gal

Background

Beta galactosidase is coded by a gene (lac z) in the lac operon of Escherichia coli. It is a metalloenzyme that splits lactose into glucose and galactose. It hydrolyzes terminal, non-reducing beta-D-galactose residues in beta-D-galactosides. Activation by cations seems to be substrate dependent. K+, Na+, NH4+, Rb+, Cs+ and Mn++ all activate enzyme activity based upon the substrate used.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • P00722(E. coli)