Key Product Details

Species Reactivity

Non-species specific

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Biotin

Antibody Source

Polyclonal Goat IgG
View all GFP Primary Antibodies »

Product Specifications

Immunogen

The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria. (Uniprot: P42212)

Reactivity Notes

No reaction was observed against Human, or Rat serum proteins. Known Cross Reactivity: rGFP. YFP differs from GFP due to a mutation at Thr203Tyr; antibodies raised against full-length GFP should also detect YFP and other variants. Reactivity in transgenic mice with GFP. Reactivity in human cell lines transfected will a GFP construct.

Specificity

No reaction was observed against Human, Mouse and Rat Serum Proteins.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room

This antibody was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, anti-biotin and purified and partially purified Green Fluorescent Protein (Aequorea victoria) Serum

Scientific Data Images for GFP Antibody [Biotin]

Western Blot Detection of GFP in HeLa Cells

Western Blot Detection of GFP in HeLa Cells

Lane 1: HeLa cells. Lane 2: mock transfected HeLa cell lysate. Load: 35 ug per lane. Primary antibody: GFP antibody at 1 ug/ml for 1 h at room temperature. Secondary antibody: IRDye 800 conjugated Donkey-a-Goat IgG [H&L] MX7 secondary antibody at 1:2,500 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 27 kDa, 33 kDa for GFP. Other band(s): none.
Immunocytochemistry/Immunofluorescence Staining of GFP in GFP-Hela Cells Using Biotin Conjugated Antibody

Immunocytochemistry/Immunofluorescence Staining of GFP in GFP-Hela Cells Using Biotin Conjugated Antibody

GFP-Biotin was tested in GFP-Hela cells against a Streptavidin conjugated 550 secondary. Hela-GFP are illuminated in Green and nuclei in Blue (DAPI).
Western Blot Detection of GFP Using Biotin Conjugated Antibody

Western Blot Detection of GFP Using Biotin Conjugated Antibody

Lane 1: GFP. Lane 2: None. Load: 50 ng per lane. Primary antibody: GFP antibody Biotin conjugated at 1:1,000 for 60 min at RT. Secondary antibody: Peroxidase streptavidin secondary antibody at 1:40,000 for 30 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 28 kDa, 28 kDa for GFP. Other band(s): None.
Immunofluorescent Staining of Mouse Cells Using Biotin Conjugated GFP Antibody

Immunofluorescent Staining of Mouse Cells Using Biotin Conjugated GFP Antibody

IF analysis of GFP in mouse aorta, liver, brain, and lung cells. Image courtesy of product review by Laura Shankman.
Immunohistochemical Detection of GFP in Free Floating Mouse Brain

Immunohistochemical Detection of GFP in Free Floating Mouse Brain

Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40um. Primary antibody: Goat anti-GFP was used at 1:500 dilution in free floating IHC to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:500 at 1:10,000 for 45 min at RT. Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.
Immunofluorescent Staining of Mouse Pancreas Using Biotin Conjugated GFP Antibody

Immunofluorescent Staining of Mouse Pancreas Using Biotin Conjugated GFP Antibody

Staining of transgenic mouse pancreas, expressing GFP in beta cells. Image courtesy of product review by Dr. Yves Heremans of Vrije Universiteit Brussel.
Immunohistochemical Staining of Drosophila Central Nervous System Tissue Using Biotin Conjugated GFP Antibody

Immunohistochemical Staining of Drosophila Central Nervous System Tissue Using Biotin Conjugated GFP Antibody

Tissue: Drosophila melanogaster late stage embryonic central nervous system. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: Anti-GFP antibody at a 1:1,000 for 1 h at RT. Secondary antibody: AlexaFluor 488 conjugated anti-Goat antibody at 1:300 for 45 min at RT. Panel A: shows a lateral view (ventral left). Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.
Detection of Purified GFP in ELISA Using Biotin Conjugated Antibody

Detection of Purified GFP in ELISA Using Biotin Conjugated Antibody

ELISA Results of GFP antibody [Biotin] tested against purified GFP protein. Each well was coated in duplicate with 10ug/mL of GFP protein. The starting dilution of antibody was 5ug/mL and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% Fish Gel/PBS Blocking buffer and SA-HRP conjugated.

Applications for GFP Antibody [Biotin]

Application
Recommended Usage

ELISA

1:50000-1:80000

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Immunohistochemistry

1:1000-1:5000

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:2000-1:10000
Application Notes
GFP Antibody is suitable for immunoblotting, ELISA, immunohistochemistry, immunomicroscopy as well as other antibody based assays using streptavidin or avidin conjugates requiring lot-to-lot consistency.

Reviewed Applications

Read 3 reviews rated 4.7 using NB100-1678 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: GFP

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

Long Name

Green Fluorescent Protein

Alternate Names

eGFP, GFPuv

Additional GFP Products

Product Documents for GFP Antibody [Biotin]

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for GFP Antibody [Biotin]

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.

Citations for GFP Antibody [Biotin]

Customer Reviews for GFP Antibody [Biotin] (3)

4.7 out of 5
3 Customer Ratings
5 Stars
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4 Stars
33%
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Transgenic mouse pancreas, expressing GFP in beta cells
    Species: Mouse
    Verified Customer | Posted 06/27/2013
    GFP Antibody [Biotin] NB100-1678
  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Mouse Aorta, Liver, Brain, Lung
    Species: Mouse
    Verified Customer | Posted 11/08/2011
    GFP Antibody [Biotin] NB100-1678
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mouse Tissue-Brain
    Species: Mouse
    Verified Customer | Posted 02/17/2010

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for GFP Antibody [Biotin]

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  • Q: We are looking for a rabbit anti-GFP antibody as a primary antibody for immunofluorescence and WB. I've seen that you carry several anti-GFP antibodies and I would like to ask which one you would recommend. The one with Cat.No NB600-308 has been used in a lot of publications and I reckon it is a solid antibody. Is this our best choice, or is there a better alternative?

    A: NB600-308 is an excellent GFP antibody and I think it would work very well for you. It has been reviewed and published with which usually makes our customer feel much more confident, seeing as it worked in other people's hand.

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