GFP tag Monoclonal antibody

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

Host / Isotype

Mouse / IgG2a

Reactivity

recombinant protein and More (5)

Applications

WB, RIP, IP, IHC, IF, FC, CoIP, ChIP, ELISA

Conjugate

Unconjugated

CloneNo.

1E10H7

Cat no : 66002-1-Ig

Synonyms

CFP, eGFP, eYFP, GFP, GFP tag, YFP



Tested Applications

Positive WB detected inGFP transgenic mouse brain tissue, Recombinant protein
Positive IP detected inTransfected HEK-293 cells
Positive IF detected inTransfected HEK-293 cells
Positive FC detected inTransfected HEK-293 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:20000-1:100000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunofluorescence (IF)IF : 1:20-1:200
Flow Cytometry (FC)FC : 0.20 ug per 10^6 cells in a 100 µl suspension
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-Ig targets GFP tag in WB, RIP, IP, IHC, IF, FC, CoIP, ChIP, ELISA applications and shows reactivity with recombinant protein samples.

Tested Reactivity recombinant protein
Cited Reactivityhuman, mouse, rat, zebrafish, pig
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 NumberU73901
Gene Symbol
Gene ID (NCBI)
RRIDAB_11182611
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC 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.



Protocols

Product Specific Protocols
WB protocol for GFP tag antibody 66002-1-IgDownload protocol
IF protocol for GFP tag antibody 66002-1-IgDownload protocol
IP protocol for GFP tag antibody 66002-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle

Nature

The CoQ oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis.

Authors - Kirill Bersuker
WB,IF

Cell Discov

Glc7/PP1 dephosphorylates histone H3T11 to regulate autophagy and telomere silencing in response to nutrient availability

Authors - Xinyu Zhang
WB

Nat Genet

Pathogenic SPTBN1 variants cause an autosomal dominant neurodevelopmental syndrome.

Authors - Margot A Cousin
IF

Cell Stem Cell

DUX-miR-344-ZMYM2-Mediated Activation of MERVL LTRs Induces a Totipotent 2C-like State.

Authors -

Nat Neurosci

Reversal of prolonged obesity-associated cerebrovascular dysfunction by inhibiting microglial Tak1.

Authors - Qing Shen
WB

Nat Biomed Eng

Restoration of dystrophin expression in mice by suppressing a nonsense mutation through the incorporation of unnatural amino acids.

Authors - Ningning Shi

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Amy (Verified Customer) (02-11-2023)

Detected EGFP-tagged constructs expressed in HEK293T cells at similar strength to other antibodies however with slightly more background.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:20,000
  • Cell Tissue Type: HEK293T
GFP tag Antibody Western Blot validation (1:20,000 dilution) in HEK293T (Cat no:66002-1-Ig)
FH

Tom (Verified Customer) (02-03-2023)

Works great. No problem. Love proteintech.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:25000
  • Cell Tissue Type: Bacteria
FH

Tatyana (Verified Customer) (01-09-2023)

Good signal, antibody can be reused. Overexpressed GFP-tagged proteins (and GFP alone) were pulled down using GFP-Trap beads and WB was done on lysates and IP samples. Antibody was diluted in 4% milk (and could be reused several times).

  • Applications: Western Blot, Immunoprecipitation
  • Primary Antibody Dilution: 1:2000
  • Cell Tissue Type: Human / HeLa cells
GFP tag Antibody Western Blot,Immunoprecipitation validation (1:2000 dilution) in Human / HeLa cells (Cat no:66002-1-Ig)
FH

Sheila (Verified Customer) (07-22-2022)

It works very well

  • Applications: Western Blot, Immunoprecipitation
  • Cell Tissue Type: Murine Breast cancer
FH

Juliana (Verified Customer) (01-28-2022)

works great for Western blot!

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:4000
  • Cell Tissue Type: HEK293T
FH

Prasanna (Verified Customer) (01-04-2021)

Of the 3 antibodies tested for GFP this one gave the cleanest and strongest signal by western blot. When I saw it on sale, I bought 8!

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:2500
  • Cell Tissue Type: Hap1, N2a (mouse neuroblastoma)
FH

Lana (Verified Customer) (12-22-2020)

SDS-PAGE: 15 ug/ul RIPA protein lysate, 4-12% Bis-Tris gradient gel.Transfer: Immobilon-FL transfer membranes (Millipore) for 2h at 80V, 4C.Blocking: SEA Block Blocking Buffer 1h, room T.Primary Ab: O/N incubation at 4C, 1:5000.Secondary Ab: IRDye 800CW Goat anti-Mouse, 1:15000.Lines of WB image: 1 – protein ladder, 2 – HEK293 whole cell lysate, negative transfection, 3 – whole cell lysate of cells transfected with eGFP.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: HEK293
GFP tag Antibody Western Blot validation (1:5000 dilution) in HEK293 (Cat no:66002-1-Ig)
FH

Thomas (Verified Customer) (11-19-2020)

HEK293T and HEK293T stably transfected with pAcGFP plasmid. 10ug total protein loaded per well. Membrane blocked 1 hour in 5% BSA prior to anti-GFP (1:2000) o/n at 4 degrees. Goat anti-mouse HRP secondary (1:10,000) used.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:2000
  • Cell Tissue Type: HEK293T
GFP tag Antibody Western Blot validation (1:2000 dilution) in HEK293T (Cat no:66002-1-Ig)
FH

Jane (Verified Customer) (03-02-2020)

Adenovirus-GFP infected cardiomyocytes stained with GFP antibody, signal is strong and effective

  • Applications: Immunohistochemistry,
  • Primary Antibody Dilution: 1:500
  • Cell Tissue Type: cardiomyocyte culture
FH

LUNFENG (Verified Customer) (01-27-2020)

GOOD

  • Applications: Immunofluorescence,
  • Cell Tissue Type: HEART
FH

Jie (Verified Customer) (01-27-2020)

Worked for western blot with GFP-LC3 transfected cardiomyocytes

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: cardiomyocytes
FH

Paul (Verified Customer) (01-15-2020)

Works well for Westerns.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:2500
  • Cell Tissue Type: HEK293T
FH

Laura (Verified Customer) (01-15-2020)

Good antibody for Western Blot.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: HEK293T
FH

Aamir (Verified Customer) (01-08-2020)

Worked well for WB

  • Applications: Western Blot,
  • Primary Antibody Dilution: 5000
  • Cell Tissue Type: HEKs
FH

Benjamin (Verified Customer) (01-07-2020)

Easily detects recombinant GFP protein via western blot with very little background.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:2500 to 1:5000
  • Cell Tissue Type: Recombinant GFP protein
FH

Jing (Verified Customer) (01-03-2020)

Used this to detect tranduce efficiency of the AAV-GFP virus. with 5% non-fat milk, there are two bands around 20-30kd, not sure which one is correct. And the antibody is relative weak, has to use 1:500 dilution.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:500
  • Cell Tissue Type: mouse heart
FH

Shan (Verified Customer) (12-25-2019)

The GFP antibody showed great sensitivity for WB and it was easily detecable. But it was insteresting that when the SDS PAGE gel separating gel concentration reeached to 12%, you can see two bands at ~30kDa and ~20kDa.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:2500
  • Cell Tissue Type: 293T
GFP tag Antibody Western Blot, validation (1:2500 dilution) in 293T (Cat no:66002-1-Ig)
FH

Jason (Verified Customer) (11-04-2019)

This is a good antibody for detecting GFP tag on Western blot, using 1:1000 dilution. The price is unbeatable, worth each penny.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: GFP-tagged protein expressed in human cell lines
FH

Hend (Verified Customer) (10-14-2019)

antibody used in western blot against egfp labelled protien and clear band was detected.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1 to 4000