Aequorea victoria GFP tag Monoclonal antibody, PBS Only

GFP tag Monoclonal Antibody for WB, IP, IF, FC, ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

recombinant protein

Applications

WB, IP, IF, FC, ELISA

Conjugate

Unconjugated

CloneNo.

1E10H7

Cat no : 66002-1-PBS

Synonyms

CFP, eGFP, eYFP, GFP, YFP

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Validation Data Gallery

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  WB analysis using 66002-1-Ig (same clone as 66002-1-PBS)

  HEK293 cells, CHO cells, normal mouse brain and GFP transgenic mouse brain were subjected to SDS PAGE followed by western blot with 66002-1-Ig (GFP tag antibody) at dilution of 1:200000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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  WB analysis of recombinant protein using 66002-1-Ig (same clone as 66002-1-PBS)

  WB result of eGFP fusion protein with anti-eGFP tag (66002-1-Ig) with GFP fusion protein at varous dilutions. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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  WB analysis of Recombinant protein using 66002-1-Ig (same clone as 66002-1-PBS)

  Recombinant protein were subjected to SDS PAGE followed by western blot with 66002-1-Ig (GFP tag antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

• 

  WB analysis of Recombinant protein using 66002-1-Ig (same clone as 66002-1-PBS)

  Recombinant protein were subjected to SDS PAGE followed by western blot with 66002-1-Ig (GFP tag antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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  IP experiment of Transfected HEK-293 using 66002-1-Ig (same clone as 66002-1-PBS)

  IP result of anti-GFP tag (IP:66002-1-Ig, 4ug; Detection:66002-1-Ig 1:2000) with the vector plasmid pEGFP-N1 Transfected HEK-293 cells lysate 300ug. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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  IF Staining of Transfected HEK-293 using 66002-1-Ig (same clone as 66002-1-PBS)

  Immunofluorescent analysis of Transfected HEK-293 cells using 66002-1-Ig (GFP tag antibody) at dilution of 1:50 and Alexa Fluor 594-conjugated AffiniPure Goat Anti-Mouse IgG (H+L). This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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  FC experiment of Transfected HEK-293 using 66002-1-Ig (same clone as 66002-1-PBS)

  1X10^6 Transfected HEK-293 cells were stained with 0.2 ug Anti-GFP tag (66002-1-Ig, Clone:1E10H7) and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), and 0.2 ug Control Antibody. Cells were fixed with 90% MeOH. This data was developed using the same antibody clone with 66002-1-PBS in a different storage buffer formulation.

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Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

66002-1-PBS targets GFP tag in WB, IP, IF, FC, ELISA 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: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS only
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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.