lacZ Rabbit Polyclonal Antibody

CAT#: R1064PS

lacZ rabbit polyclonal antibody, Purified

Product Images

Western blot using Beta-galactosidase antibody. shows detection of a band at ~117 kDa (lane 1) corresponding to the protein present in partially purified preparations. Approximately 50 ng of protein was separated on a 4-20% Tris-Glycine gel by SDSPAGE 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 IRDye800 (TM) conjugated goat-a-rabbit IgG [H&L] for 45 min at RT (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). RDye800 (TM) fluorescence image was captured using the Odyssey (R) infrared imaging system developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.

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  Western blotting using Beta-galactosidase antibody. Lane 1 shows 80 ng and lane 2 shows 20 ng loaded onto gel. Results for N<em>on-Reducing Conditions</em> of SDS-PAGE prior to transfer to nitrocellulose are shown on the left side of the figure; results obtainined under <em>Reducing Conditions</em> are shown on the right. Blots were blocked overnight at 4°C with blocking buffer for fluorescent Western blotting (p/n MB-070). The membrane was probed with Beta-galactosidase antibody [R1064P] diluted to 1/10,000. Reaction occurred overnight at 4°C. Dylight649?™ conjugated goat-anti-rabbit IgG was used for detection. Molecular weight estimation was made by comparison to a prestained MW marker (center). Fluorescence image was captured using the VersaDoc® imaging system developed by BIO-RAD. Other detection systems will yield similar results.

Specifications

Product Data

• Applications: ELISA, IF, IHC, IP, WB
• Recommended Dilution: Suitable for Immunoblotting (Western or dot blot), ELISA, Immunofluorescence microscopy, Immunoprecipitation, conjugation and most immunological methods requiring high titer and specificity.
  Western blot: 1/5,000-1/10,000. A 1/5,000 dilution has been reported to be successful for staining by Immunoblot of Beta-galactosidase fusion proteins after transfer using a semi-dry transfer apparatus). ELISA: 1/10,000.
  Immunohistochemistry: 1/1,500. The antibody recognizes both Frozen tissue sections, Paraffin embedded tissue and 4% paraformaldehyde fixed tissue for most immunohistochemical analysis:A 1/1,500 dilution has been reported to detect Beta-galactosidase in adult rat spinal cord tissue after infection with helper-dependent adenovirus expressing lacZ. In this particular experiment, tissue was perfused with 4% paraformaldehyde and cryostat-cut (35 µm) to produce free-floating sections. A 1/5,000 dilution has been reported for immunofluorescent staining of methanol fixed, devitellinized Drosophila embryos. Although a wide range of conditions was reported to be effective, a 1/10,000 dilution was noted to show no background and to be suitable for double labeling experiments). A 1/5,000 dilution has been reported to be successful for staining brain sections from transgenic mice expressing nuclear Beta-galactosidase when assayed by Immunofluorescence microscopy.
• Reactivities: Escherichia coli
• Host: Rabbit
• Clonality: Polyclonal
• Immunogen: Full length native Beta-galactosidase isolated from E.coli
• Specificity: This antibody detects Beta-galactosidase [E. coli]. Cross reactivity against Beta-galactosidase from other tissues and species may occur but have not been specifically determined. Very low background staining has been reported in various assays.
  Immunoelectrophoresis gives a single precipitin arc against anti-rabbit serum as well as purified and partially purified Beta-galactosidase [E.coli].
• Formulation: 0.02M Potassium phosphate, 0.15M Sodium chloride, pH 7.2
  State: Purified
  State: Lyophilized purified IgG fraction
  Stabilizer: None
  Preservative: 0.01% (w/v) Sodium azide
• Reconstitution Method: Restore with 0.1 ml of deionized water (or equivalent).
• Concentration: lot specific
• Purification: Multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer
• Conjugation: Unconjugated
• Storage: Store lyophilized at 2-8°C for 6 months or at -20°C long term.
  After reconstitution store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C long term. Dilute only prior to immediate use. Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Database Link:
  UniProt ID P00722
• 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.
  Anti Beta-galactosidase antibody recognizes the enzyme beta-galactosidase, or ß-galactosidase, that is a component of assays used frequently in genetics, molecular biology (see X-gal) for a blue white screen, and other life sciences. IPTG induces production of ß-galactosidase by binding and inhibiting the lac repressor. Since it is highly expressed and accumulated in lysosomes in senescent cells, it is used as a senescence biomarker both in vivo and in vitro in qualitative and quantitative assays, despite its limitations. Anti Beta-galactosidase antibody is ideal for investigators involved in enzyme research.
• Synonyms: Beta-Gal tag, lacZ tag, b0344, JW0335, Beta-Gal Fusion Protein, Lactase
• Note: Conjugates available. Please ask for details.