EGFP | Enhanced Green Fluorescence Protein

AS21 4583 | Clonality: Polyclonal | Host: Rabbit | Reactivity: EGFP

EGFP | Enhanced Green Fluorescence Protein

Product Information

Immunogen

Recombinant GFP from Aequorea coerulescens (belt jellyfish) overexpressed and purified from E.coli, UniProt: Q6YGZ0

Host Rabbit

Clonality Polyclonal

Purity Immunogen affinity purified in PBS pH 7.2-7.6, 0.1% trehalose, 5 mM sodium azide.

Format Lyophilized

Quantity 100 µg

Reconstitution For reconstitution add 100 µl of sterile water, ultrapure water

Storage Store lypholized antibody at 4 ºC. After reconstitution keep aliquots at -20ºC for a higher stability. Avoid repetitive freeze/thaw cycles. Centrifuge briefly to remove any insoluble material.Expiry date: 12 months after purchase if unopened

Tested applications Immunocytochemistry (ICC), Western blot (WB)

Recommended dilution 1:2000 - 1:5000 (ICC), 1:1000 - 1:5000 (WB)

Expected | apparent MW 27 kDa (for GFP without fusion partner)

Reactivity

Confirmed reactivity GFP, EGFP

Not reactive in mCherry

Application examples

Application examples Immunolocalization and western blot using anti-GFP rabbit polöyclonal antibodies

Left panel: Analysis of GFP expression (green) in transfected HEK293 cells by immunocytochemistry. Cells were stained with rabbit anti-GFP antibody (red, 1:2,000). Blue: Hoechst staining for nuclei. Transfected cells appear orange-yellow due to the green (GFP) and red (anti-GFP antibody) color overlay.
Right panel: Western blot analysis of GFP protein in transfected HEK293 lysate using rabbit antibody to GFP (green, 1:2,000). [1] protein standard, [2] non-transfected control cells, [3] transfected cells with GFP construct, [4] transfected cells with mCherry construct. A strong band at ~27 kDa corresponds to the expected molecular weight of GFP protein detected only in cells transfected with GFP construct. The antibody does not cross-react with mCherry protein. The same blot was simultaneously probed with a mouse antibody to beta-tubulin (red, ~ 50 kDa, lanes 2-4).

Additional information

EGFP has Excitation max (nm): 488; Emission max (nm): 509; Extinction coefficient (Cm-1M-1): 61000

Background

Background EGFP is an Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Xaa-N and Gly-(N+2), and oxidation of Tyr-(N+1) to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

Product citations

calculated \| apparent molecular mass [kDa]:	27 kDa (for GFP without fusion partner)
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Rabbit
immunogen:	Recombinant GFP from Aequorea coerulescens (belt jellyfish) overexpressed and purified from E.coli, UniProt: Q6YGZ0
Purity:	Immunogen affinity purified in PBS pH 7.2-7.6, 0.1% trehalose, 5 mM sodium azide.
Quantity:	100 µg
recommended dilution:	1:2000 - 1:5000 (ICC), 1:1000 - 1:5000 (WB)
Reconstitution:	For reconstitution add 100 µl of sterile water, ultrapure water
storage:	Store lypholized antibody at 4 ºC. After reconstitution keep aliquots at -20ºC for a higher stability. Avoid repetitive freeze/thaw cycles. Centrifuge briefly to remove any insoluble material.Expiry date: 12 months after purchase if unopened
tested applications:	Immunocytochemistry (ICC), Western blot (WB)
Picture (footer):	Left panel: Analysis of GFP expression (green) in transfected HEK293 cells by immunocytochemistry. Cells were stained with rabbit anti-GFP antibody (red, 1:2,000). Blue: Hoechst staining for nuclei. Transfected cells appear orange-yellow due to the green (GFP) and red (anti-GFP antibody) color overlay. Right panel: Western blot analysis of GFP protein in transfected HEK293 lysate using rabbit antibody to GFP (green, 1:2,000). [1] protein standard, [2] non-transfected control cells, [3] transfected cells with GFP construct, [4] transfected cells with mCherry construct. A strong band at ~27 kDa corresponds to the expected molecular weight of GFP protein detected only in cells transfected with GFP construct. The antibody does not cross-react with mCherry protein. The same blot was simultaneously probed with a mouse antibody to beta-tubulin (red, ~ 50 kDa, lanes 2-4).
additional information (application):	EGFP has Excitation max (nm): 488; Emission max (nm): 509; Extinction coefficient (Cm-1M-1): 61000
background:	EGFP is an Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Xaa-N and Gly-(N+2), and oxidation of Tyr-(N+1) to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.
Confirmed reactivity:	GFP, EGFP
not reactive in:	mCherry

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