Figure 1. Flow cytometric analysis of GFP expression in transfected human embryonic kidney cells. AcGFP-transfected HEK-293 cells were fixed with BD Cytofix™ Fixation Buffer (554655) and permeabilized with the BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-GFP monoclonal antibody (solid line, Cat. No. 565197). The histograms were derived from gated events based on light scattering characteristics of intact HEK-293 cells. Flow cytometry was performed on a BD LSRFortessa™ Cell Analyzer System. Figure 2. Immunofluorescent staining of GFP-transfected human embryonic kidney cells. Untransfected HEK-293 cells (left) and AcGFP-transfected HEK-293 cells (middle, right) were fixed with BD Cytofix™ Fixation Buffer (554655) and permeabilized with the BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with Alexa Fluor® 647 Mouse Anti-GFP monoclonal antibody (pseudo-colored red) at 20 μg/mL. GFP signal in the AcGFP-transfected HEK-293 cells is shown for comparison (pseudo-colored green). Cell nuclei were counterstained with BD Pharmingen™ DAPI (pseudo-colored blue, Cat. No. 564907). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.
Figure 1. Flow cytometric analysis of GFP expression in transfected human embryonic kidney cells. AcGFP-transfected HEK-293 cells were fixed with BD Cytofix™ Fixation Buffer (554655) and permeabilized with the BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-GFP monoclonal antibody (solid line, Cat. No. 565197). The histograms were derived from gated events based on light scattering characteristics of intact HEK-293 cells. Flow cytometry was performed on a BD LSRFortessa™ Cell Analyzer System. Figure 2. Immunofluorescent staining of GFP-transfected human embryonic kidney cells. Untransfected HEK-293 cells (left) and AcGFP-transfected HEK-293 cells (middle, right) were fixed with BD Cytofix™ Fixation Buffer (554655) and permeabilized with the BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with Alexa Fluor® 647 Mouse Anti-GFP monoclonal antibody (pseudo-colored red) at 20 μg/mL. GFP signal in the AcGFP-transfected HEK-293 cells is shown for comparison (pseudo-colored green). Cell nuclei were counterstained with BD Pharmingen™ DAPI (pseudo-colored blue, Cat. No. 564907). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.
製品詳細
BD Pharmingen™
Mouse IgG1, κ
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development), Western blot (Not Recommended)
0.2 mg/ml
AB_2739107
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- 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.
関連製品
Fixation Buffer RUO
サイズ
100 mL
カタログ番号
554655
Perm Buffer III RUO
サイズ
125 mL
カタログ番号
558050
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
サイズ
100 Tests
カタログ番号
557714
DAPI Solution RUO
サイズ
1 mg
カタログ番号
564907
565197 Rev. 1
抗体の詳細
1A12-6-18
Monoclonal antibody 1A12-6-18 reacts with Green Fluorescent Protein (GFP), which is a 27 kDa bioluminescent protein first purified from the jellyfish, Aequorea victoria. GFP exhibits green fluorescence when exposed to blue or ultraviolet light. Additionally, GFP can be introduced into the genome of and expressed in a wide variety of cell types, making it a useful reporter for tracking gene expression or protein localization. A number of GFP mutants have been developed with varying fluorescence intensity and spectra. Monoclonal antibody 1A12-6-18 is known to react with AcGFP, and is not reactive towards ZsGreen1.
565197 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 520-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
565197 Rev.1
引用＆参考文献
View product citations for antibody "565197" on CiteAb
Development References (4)
-
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994; 263(5148):802-805. (Biology). View Reference
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Heinen AP, Wanke F, Moos S, et al. Improved Method to Retain Cytosolic Reporter Protein Fluorescence While Staining for Nuclear Proteins. Cytometry A. 2014; 85A:621-627. (Biology). View Reference
-
Kain SR, Adams M, Kondepudi A, Yang TT, Ward WW, Kitts P. Green fluorescent protein as a reporter of gene expression and protein localization. Biotechniques. 1995; 19(4):650-655. (Biology). View Reference
-
Nybo K. GFP Imaging in Fixed Cells. Biotechniques. 2012; 52(6):359-360. (Biology). View Reference
565197 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.