CoraLite®594-conjugated GFP tag Monoclonal antibody

GFP tag Monoclonal Antibody for FC (Intra), IF, WB

Host / Isotype

Mouse / IgG2a

Reactivity

recombinant protein

Applications

WB, IF, FC (Intra)

Conjugate

CoraLite®594 Fluorescent Dye

CloneNo.

1E10H7

Cat no : CL594-66002

Synonyms

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Validation Data Gallery

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  WB analysis using CL594-66002

  Recombinant protein were subjected to SDS PAGE followed by western blot with CL594-66002 (GFP tag antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IF Staining of Transfected HEK-293 using CL594-66002

  Immunofluorescent analysis of (-20°C Ethanol) fixed Transfected HEK-293 cells using CoraLite®594-conjugated GFP tag antibody (CL594-66002, Clone: 1E10H7 ) at dilution of 1:200.

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  FC experiment of Transfected HEK-293 using CL594-66002

  1X10^6 Transfected HEK-293 cells were stained with 0.2 ug CoraLite®594-conjugated Anti-GFP tag (CL594-66002, Clone:1E10H7) (red), and 0.2 ug Control Antibody. Cells were fixed with 90% MeOH.

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Tested Applications

• Positive WB detected in: Recombinant protein
• Positive IF detected in: Transfected HEK-293 cells
• Positive FC detected in: Transfected HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Product Information

CL594-66002 targets GFP tag in WB, IF, FC (Intra) applications and shows reactivity with recombinant protein samples.

• Tested Reactivity: recombinant protein
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: GFP tag fusion protein Ag2128
• Full Name: GFP tag
• Calculated molecular weight: 26 kDa
• GenBank accession number: U73901
• Conjugate: CoraLite®594 Fluorescent Dye
• Excitation/Emission maxima wavelengths: 588 nm / 604 nm
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Green fluorescence protein (GFP) is a protein composed of 238 amino acid residues (26.9kDa) derived from the jellyfish Aequorea Victoria which emits green light (emission peak at 509nm) when excited by blue light (excitation peak at 395nm). GFP, when exposed to light in the blue to ultraviolet spectrum, will show a bright green fluorescent light, making it a very useful tool in research.

What is the molecular weight of GFP?

26.9 kDa

How does GFP work?

GFP was first isolated from the jellyfish Aequorea Victoria, a source of bioluminescence, in the 1960s and in 2008 the Nobel Prize in Chemistry was awarded "for the discovery and development of the green fluorescent protein, GFP" to Osamu Shimomura and colleagues, who recognized its potential in research (PMID: 13911999). A short amino acid sequence within the protein acts as the chromophore, which absorbs UV light at 395 nm and emits green light at 509 nm. 

Why is GFP a useful reporter?

When GFP was sequenced in 1992 (PMID: 1347277) it allowed scientists to express it in other organisms using transgenic techniques. It does not require cofactors to work, is non-toxic to live cells, and is relatively small, making it ideal as a "tag" for other proteins, identifiable by shining a UV light and observing the green fluorescence. The tertiary folded structure of GFP forms a chromophore at the center of a barrel shape, which protects the fluorescence-emitting amino acid chain from solvents, meaning it can function in many environments (PMID 9759496).

What are the applications for GFP?

When expressed attached to another protein, GFP can be used as a reporter gene to measure expression levels or can easily be used in fluorescence microscopy. It has been used to highlight proteins in a variety of model organisms, including bacteria, zebrafish, and mice.