Flow cytometric analysis of β-galactosidase expression in Escherichia coli (strain K12) β-galactosidase-transfected cells. Non-transfected cells (Left Plot) or β-galactosidase-transfected cells (Right Plot) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with Perm/Wash Buffer (Cat. No. 554723). The cells were then stained with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; dashed line histogram) or Alexa Fluor™ 647 Rat Anti-β-galactosidase antibody (Cat. No. 570540/570541; solid line histogram) at 0.25 µg/test. The fluorescence histogram showing β-galactosidase expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of β-galactosidase expression in Escherichia coli (strain K12) β-galactosidase-transfected cells. Non-transfected cells (Left Plot) or β-galactosidase-transfected cells (Right Plot) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with Perm/Wash Buffer (Cat. No. 554723). The cells were then stained with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; dashed line histogram) or Alexa Fluor™ 647 Rat Anti-β-galactosidase antibody (Cat. No. 570540/570541; solid line histogram) at 0.25 µg/test. The fluorescence histogram showing β-galactosidase expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
製品詳細
BD Pharmingen™
beta-gal; beta-galactosidase; lacZ; lactase; β-gal; β-galactosidase
Bacterial (QC Testing)
Rat IgG1, κ
E. coli β-galactosidase
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
945006
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
推奨アッセイ手順
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
関連製品
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
Fixation Buffer RUO
サイズ
100 mL
カタログ番号
554655
Perm/Wash Buffer RUO
サイズ
100 mL
カタログ番号
554723
Alexa Fluor® 647 Rat IgG1, κ Isotype Control RUO
サイズ
0.1 mg
カタログ番号
557731
570541 Rev. 1
抗体の詳細
13R4.rMAb
13R4.rMAb is a recombinant monoclonal antibody that specifically recognizes Escherichia coli β-galactosidase, which is also known as beta-galactosidase or beta-gal. β-galactosidase is a cytoplasmatic metalloenzyme encoded by the lacZ gene in the lac operon of E. coli. β-galactosidase catalyzes the hydrolysis of lactose to its constituent monosaccharides glucose and galactose. E. coli β-galactosidase is often used as a tag for recombinant mammalian proteins because it is not expressed by mammalian cells. This permits easy detection and purification of β-galactosidase-tagged recombinant proteins expressed by genetically engineered mammalian cells.
570541 Rev. 1
フォーマットの詳細
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
570541 Rev.1
引用＆参考文献
Development References (5)
-
Juers DH, Matthews BW, Huber RE. LacZ β-galactosidase: structure and function of an enzyme of historical and molecular biological importance.. Protein Sci. 2012; 21(12):1792-807. (Biology). View Reference
-
Laden JC, Philibert P, Torreilles F, Pugnière M, Martineau P. Expression and folding of an antibody fragment selected in vivo for high expression levels in Escherichia coli cytoplasm.. Res Microbiol. 2002; 153(7):469-74. (Biology). View Reference
-
Martineau P, Jones P, Winter G. Expression of an antibody fragment at high levels in the bacterial cytoplasm.. J Mol Biol. 1998; 280(1):117-27. (Immunogen: ELISA, Western blot). View Reference
-
Philibert P, Martineau P. Directed evolution of single-chain Fv for cytoplasmic expression using the beta-galactosidase complementation assay results in proteins highly susceptible to protease degradation and aggregation.. Microb Cell Fact. 2004; 3(1):16. (Clone-specific: Functional assay). View Reference
-
Sibler AP, Courtête J, Muller CD, Zeder-Lutz G, Weiss E. Extended half-life upon binding of destabilized intrabodies allows specific detection of antigen in mammalian cells.. FEBS J. 2005; 272(11):2878-91. (Clone-specific: Fluorescence activated cell sorting, Intracellular Staining/Flow Cytometry, Western blot). View Reference
570541 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
