GFP Antibody

Images

 
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Staining of cultured hippocampal neuron from rat. ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - HeLa cells transiently expressing GFP were visualized using GFP antibody (green). Nuclei were counterstained with DAPI. ICC/IF image submitted by a ...read more
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - IF analysis of GFP in mouse trigeminal ganglia, cornea. Image courtesy of product review by Jessica Newsom.
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Comparison between GFP immunoreactivity in Rexed Lamina 2 neurons of a transgenic mouse using anti-GFP (GREEN) and rabbit anti-GFP (RED). The ...read more
Immunohistochemistry: GFP Antibody [NB100-1614] - 5dpf Tg(brn3c:GFP) larvae were incubated with vehicle alone (DMSO, A), 400uM of cisplatin (CP, B), pre-treated with Qx for 2 hours and then incubated with CP for 6 hours ...read more
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - IF analysis of GFP in rat cortical culture (2 weeks old). ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - LNCaP human prostate cancer cell line imaged after transfection with GFP expressing plasmid. ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Staining of a tissue section through the midbrain region of an adult mouse for GFP (green) and Tryptophan Hydroxylase-positive neurons (RED).
Immunohistochemistry-Paraffin: GFP Antibody [NB100-1614] - Pyramidal neurons in the hippocampal formation of a neonatal mouse brain. Tissue was paraformaldehyde-fixed (4%) and paraffin-embedded. GFP staining is in green ...read more
Immunohistochemistry-Paraffin: GFP Antibody [NB100-1614] - Embyonic mouse brain section. GFP in green, DAPI in grey. IHC-P image submitted by a verified customer review.
Immunohistochemistry: GFP Antibody [NB100-1614] - (J) control. (K) Neo 200uM 30min. (L) GM 50uM 1 hour. (M) CP: 400uM 2 hours. (N) Qx 300uM 8 hours. (O) Qx 300uM 8 hours + Neo 200uM 30min. (P) Qx 300uM 8 hours + GM 50uM ...read more

Product Details

Summary
Reactivity Mu, All-NA, DrSpecies Glossary
Applications WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Chicken
Conjugate
Unconjugated
Concentration
10.0 mg/ml

Order Details

GFP Antibody Summary

Immunogen
This GFP antibody was developed by immunizing chickens with purified recombinant green fluorescent protein (GFP) emulsified in Freund's adjuvant.
Isotype
IgY
Clonality
Polyclonal
Host
Chicken
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:5000 - 1:10000
  • ELISA 1:100 - 1:2000
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry 1:2000 - 1:5000
  • Immunohistochemistry-Frozen 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
Application Notes
Antibodies were analyzed by Western blot analysis (1:5000 dilution) and immunohistochemistry (1:500 dilution) using transgenic mice expressing the GFP gene product. Western blots were performed using BlokHen as the blocking reagent, and HRP-labeled goat anti-chicken antibodies as the detection reagent. Immunohistochemistry used tetramethyl rhodamine-labeled anti-chicken IgY. Use in IHC-F was reported in the scientific literature (PMID: 23799397).
Reviewed Applications
Read 6 Reviews rated 4.3
using
NB100-1614 in the following applications:

Publications
Read Publications using
NB100-1614 in the following applications:

Reactivity Notes

NB100-1614 has been tested on transgenic mice expressing recombinant GFP. Mouse reactivity reported in multiple pieces of scientific literature. Drosophila reactivity reported in scientific literature (PMID: 28192468).

Packaging, Storage & Formulations

Storage
Store at -20C in the dark. Avoid freeze-thaw cycles.
Buffer
10 mM PBS (0.9% isotonic, w/v, pH 7.2) and 50% (v/v) Glycerol
Preservative
0.02% Sodium Azide
Concentration
10.0 mg/ml
Purity
Immunogen affinity purified

Notes

Chicken products cannot be exported to Canada.

Purification Notes
Chickens were immunized with purified recombinant green fluorescent protein (GFP) emulsified in Freund's adjuvant. After multiple injections, eggs were collected from the hens, and IgY fractions were prepared from the yolks and then affinity-purified antibodies were prepared using GFP conjugated to an agarose matrix. The final product is a filter-sterilized mixture of both affinity-purified antibodies (30 ug/ml) and purified IgY (10 mg/mL).

Storage Notes
Store at -20C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Since 50% glycerol is present in the vial, this antibody preparation should remain a liquid at -20C. For longer storage periods, store at -80C, but be aware that freezing this preparation may reduce its activity.

Alternate Names for GFP Antibody

  • eGFP
  • Enhanced Green Fluorescent Protein
  • GFP
  • GFPuv
  • green fluorescent protein (gfp)
  • Green Fluorescent Protein

Background

Green fluorescent protein (GFP) is a 27 kDa protein derived from the bioluminescent jellyfish Aquorea Victoria. GFP can be excited by light in the blue to ultraviolet spectrum and has an excitation peak at 395 nm with an emission peak at 509 nm in the green spectrum. GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. GFP fluorescence is stable under fixation conditions and antibodies are suitable for a variety of applications including ELISA, ICC/IF, IHC, IP and WB (1-3).

References
1. Shi, C., Pan, F. C., Kim, J. N., Washington, M. K., Padmanabhan, C., Meyer, C. T.,... Means, A. L. (2019). Differential Cell Susceptibilities to Kras(G12D) in the Setting of Obstructive Chronic Pancreatitis. Cell Mol Gastroenterol Hepatol. doi:10.1016/j.jcmgh.2019.07.001

2. Zhao, S., Fortier, T. M., & Baehrecke, E. H. (2018). Autophagy Promotes Tumor-like Stem Cell Niche Occupancy. Curr Biol, 28(19), 3056-3064.e3053. doi:10.1016/j.cub.2018.07.075

3. Zusso, M., Lunardi, V., Franceschini, D., Pagetta, A., Lo, R., Stifani, S.,... Moro, S. (2019). Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway. J Neuroinflammation, 16(1), 148. doi:10.1186/s12974-019-1538-9

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GFP Antibody (NB100-1614)(39)

We have publications tested in 9 confirmed species: Human, Mouse, Rat, Drosophila, Firefly, Jellyfish, Primate, Yeast, Zebrafish.

We have publications tested in 7 applications: ICC/IF, IHC, IHC-Fr, IHC-FrFl, IHC-P, IHC/IF, WB.


Filter By Application
ICC/IF
(10)
IHC
(14)
IHC-Fr
(4)
IHC-FrFl
(1)
IHC-P
(3)
IHC/IF
(1)
WB
(1)
All Applications
Filter By Species
Human
(1)
Mouse
(13)
Rat
(1)
Drosophila
(1)
Firefly
(1)
Jellyfish
(2)
Primate
(1)
Yeast
(1)
Zebrafish
(2)
All Species
Showing Publications 1 - 10 of 39. Show All 39 Publications.
Publications using NB100-1614 Applications Species
Tian W, Qin S, Zhang B et Al. T606-phosphorylated Kaiso interacting with 14-3-3 and p120ctn is sequestered in the cytoplasm of human cells bioRxiv Jan 1 2020 (WB, Human) WB Human
Perniss A, Schmidt P, Soultanova A et al. Development of epithelial cholinergic chemosensory cells of the urethra and trachea of mice Cell and tissue research Feb 22 2021 [PMID: 33616728] (Mouse) Mouse
Chiola S, Napan KL, Wang Y et al. Defective AMPA-mediated synaptic transmission and morphology in human neurons with hemizygous SHANK3 deletion engrafted in mouse prefrontal cortex Molecular psychiatry Feb 8 2021 [PMID: 33558651] (IHC-FrFl, Mouse) IHC-FrFl Mouse
Hollon N, Williams E, Howard C, et al. Nigrostriatal Dopamine Signals Sequence-Specific Action-Outcome Prediction Errors bioRxiv Jan 26 2021 (IHC) IHC
Zallocchi M, Hati S, Xu Z, et al. Characterization of quinoxaline derivatives for protection against iatrogenic-induced hearing loss JCI insight Jan 21 2021 [PMID: 33476306] (ICC/IF) ICC/IF
Bailly J, Del Rossi N et al. Targeting Morphine-Responsive Neurons: Generation of a Knock-In Mouse Line Expressing Cre Recombinase from the Mu-Opioid Receptor Gene Locus. Eneuro Oct 5 2020 [PMID: 32381649] (IHC) IHC
Xu J, Wang L, Li H et al. Short stature homeobox 2 (SHOX2) regulates osteogenic differentiation and pattern formation during hard palate development in mice J. Biol. Chem. 2019 Oct 24 [PMID: 31649032] (IHC-P, IHC, Mouse) IHC-P, IHC Mouse
Madsen SD, Jones SH, Tucker HA et al. Survival of aging CD264+ and CD264- populations of human bone marrow mesenchymal stem cells is independent of colony-forming efficiency Biotechnol. Bioeng. Oct 15 2019 [PMID: 31612990]
Aoki S, Smith JB, Li H et al. An open cortico-basal ganglia loop allows limbic control over motor output via the nigrothalamic pathway Elife Sep 6 2019 [PMID: 31490123] (IHC/IF, Rat) IHC/IF Rat
Ehrlich A T, Semache M et al. Biased Signaling of the Mu Opioid Receptor Revealed in Native Neurons. iScience Mar 15 2019 [PMID: 30925410] (ICC/IF) ICC/IF
Show All 39 Publications.

Reviews for GFP Antibody (NB100-1614) (6) 4.36

Average Rating: 4.3
(Based on 6 reviews)
We have 6 reviews tested in 3 species: Human, Mouse, Rat.

Reviews using NB100-1614:
Filter by Applications
IHC-P
(1)
ICC
(2)
IF
(2)
IHC
(1)
All Applications
Filter by Species
Human
(2)
Mouse
(2)
Rat
(2)
All Species
Images Ratings Applications Species Date Details
Immunohistochemistry-Paraffin GFP NB100-1614
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4
reviewed by:
Olivier Etienne
IHC-P Mouse 02/17/2020
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedembryonic mouse brain
SpeciesMouse
LotGFP879484
Immunocytochemistry GFP NB100-1614
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4
reviewed by:
Anonymous
ICC Human 05/09/2018
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Summary

ApplicationImmunocytochemistry
Sample TestedLNCaP human prostate cancer cell line
SpeciesHuman
LotB-2

Comments

CommentsHuman LnCap cells were grown in RPMI1640 + 10% FCS in 5% CO2 at 37C. The cells were electroporated with GFP expressing plasmid using Amaxa Nucleofection kit.
Immunocytochemistry GFP NB100-1614
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5
reviewed by:
Anonymous
ICC Human 12/25/2016
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Summary

ApplicationImmunocytochemistry
Sample Tested hela cell
SpeciesHuman

Comments

CommentsGFP expressing cells were fixed with PFA 4% for 30 min, washed several times with PBS and then permeabilized for 5 min with PBSTx-100 1%. After several washes cells were incubated with the chicken anti-GFP 1:500 in fish blocking solution (PBS + 2.5% fish gelatin + 2.5% FCS) overnight at 4C. The next day after several washes with PBS, cells were incubated with anti-chicken Alexa 488 1:400 in fish gelatin for 1hour at room temperature.
Immunofluorescence GFP NB100-1614
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4
reviewed by:
Anonymous
IF Rat 02/17/2016
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Summary

ApplicationImmunofluorescence
Sample Testedrat cortical culture (2 weeks old)
SpeciesRat

Blocking

Blocking Details4% BSA, 4%goat serum, 0.1%TritonX-100 in TBS. Blocking for 1hr at 4C

Primary Anitbody

Dilution Ratio1:500 for surface GFP labeling, 1:1000 for total GFP labeling

Secondary Antibody

Secondary Descriptiongoat anti-chicken Alexa fluor 488, goat anti-chicken Alexa fluor 568
Secondary Manufacturer Cat#ThermoScientific: A-11039, A-11041
Secondary Concentration0.4mg/mL (1:500 dilution)

Details

Detection NotesZeiss confocal microscopy 710 (63x oil objective, pixel time 0.64us )
Fixation DetailsLive neurons were fed with GFP antibody in culture media for 10min at 37C before fixation in fixation buffer(4%PFA,4%sucrose TBS
Wash DescriptionTBS. Wash 3 times (5min each) between each steps.

Comments

CommentsI aimed to perform both surface and total GFP-GluA2 labeling in neurons. Here is a brief summary of the protocal. For surface labeling, the live neurons were fed with GFP antibodies for 10min followed by fixation with 4% PFA for 15min. Then sample were blocked in non-permeable blocking buffer (without TritonX-100)for 1hr, followed by goat anti chicken Alexa Fluor 488. Then the sample were blocked in permeable blocking buffer (with TritonX-100) for 1hr or overnight before incubation with GFP primary antibody followed by goat anti-chicken Alexa fluor 568. After final wash, the samples were mounted to glass slide with ProLong® Diamond Antifade Mountant (Thermoscitific P36961).
Immunofluorescence GFP NB100-1614
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5
reviewed by:
Hongtian Yang
IF Rat 08/23/2013
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Summary

ApplicationImmunofluorescence
Sample Testedcultured hippocampal neuron from rat
SpeciesRat
Lot0511FP12
CommentsThe chicken GFP antibody worked very well. The reason we choose chicken the version of Novus' GFP antibody is because it allows us to do double or triple staining if necessary, since most other primary antibody's are usually from rabbit or mouse.

Blocking

Blocking Details0.1% TritonX-100, 5% goat serum in PBS; 1hr, 4C

Primary Anitbody

Dilution Ratio1:800, overnight, 4C

Secondary Antibody

Secondary Descriptiongoat anti-chicken IgG (H+L), Alexa 488
Secondary Manufacturer Cat#A11039
Secondary Concentration4ug/ml

Details

Detection NotesZeiss Confocal Microscopy
Fixation Details4% formaldehyde, PBS, room temperture
Wash DescriptionPBS, 10min, 3 times per solution change

Comments

CommentsThe chicken GFP antibody worked very well. The reason we choose chicken the version of Novus' GFP antibody is because it allows us to do double or triple staining if necessary, since most other primary antibody's are usually from rabbit or mouse.
Immunohistochemistry GFP NB100-1614
Enlarge
4
reviewed by:
Anonymous
IHC Mouse 05/15/2012
View

Summary

ApplicationImmunohistochemistry
Sample TestedMouse Trigeminal Ganglia, Cornea
SpeciesMouse
Lot0316FP11
CommentsUsed 1:40 dilution of primary at 1:8000

Blocking

Blocking Details5% NGS

Primary Anitbody

Dilution Ratio1:40 dilution in NGS/PBS, overnight incubation at room temperature

Secondary Antibody

Secondary DescriptionGoat anti-chicken
Secondary Manufacturer Cat#A11039
Secondary Concentration1:10 dilution in 10% NGS, used at 1:1500

Details

Detection NotesCut tissue via cryostat, viewed on Nikon E800 microscope at 10, 20x. Exposure time was negligible. Only negative controls used
Fixation Details20um sections of formalin-fixed tissue
Wash DescriptionWash 3x for 10min each in 1%NGS

Comments

CommentsUsed 1:40 dilution of primary at 1:8000

Product General Protocols

View specific protocols for GFP Antibody (NB100-1614): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for GFP Antibody (NB100-1614). (Showing 1 - of FAQs).

    Secondary Antibodies

     

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    GFP, or green fluorescent protein, is a chemiluminescent protein derived from Aequorea jellyfish that was first discovered by Osamu Shimomura.  It was soon after established that the emission spectra of GFP was right around 509nm, or the ultraviol...  Read full blog post.

    GFP - Be Green!
    Green fluorescence protein (GFP) is a 27KD protein derived from the jellyfish Aquorea victoria that emits a green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). GFP is a highly v...  Read full blog post.

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    Recent Reviews

    4.3
    5
    2
    4
    4
    3
    0
    2
    0
    1
    0

    Olivier Etienne
    02/17/2020
    Application: IHC-P
    Species: Mouse

    Anonymous
    05/09/2018
    Application: ICC
    Species: Human

    Anonymous
    12/25/2016
    Application: ICC
    Species: Human