Key Product Details

Species Reactivity

Non-species specific

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Knockdown Validated

Cited:

Western Blot

Label

HRP

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-Peroxidase and purified and partially purified Green Fluorescent Protein (Aequorea victoria)

Scientific Data Images for GFP Antibody [HRP]

Western Blot Detection of GFP Expressed in HEK293 Cells Using HRP Conjugated Antibody

Western Blot Detection of GFP Expressed in HEK293 Cells Using HRP Conjugated Antibody

This GFP antibody works well with mGFP tagged over-expression. Because it is conjugated with HRP, secondary antibody is not needed, saves lots of time. Image from verified customer review.
Western Blot Detection of GFP Using HRP Conjugated Antibody

Western Blot Detection of GFP Using HRP Conjugated Antibody

Lane 1: 50ng of GFP. Lane 2: none. Primary antibody: none. Secondary antibody: Anti-GFP Antibody Peroxidase Conjugate secondary antibody was used at 1:1000 for 45 min at RT. Block: 1% BSA-TTBS 30 min at 20C. Predicted/Observed size: 28 kDa for GFP. Other band(s): none
Use of HRP Conjugated GFP Antibody in Western Blot

Use of HRP Conjugated GFP Antibody in Western Blot

Western Blot of Goat anti-GFP antibody Peroxidase conjugated. Marker: Opal Pre-stained ladder

Applications for GFP Antibody [HRP]

Application
Recommended Usage

ELISA

1:10000-1:50000

Immunohistochemistry

1:500-1:2000

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:2000-1:5000
Application Notes
This product is designed to detect GFP and its variants. Anti-GFP Peroxidase conjugated antibody has been tested by ELISA to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP and by western blot. Biotin conjugated polyclonal anti-GFP used in a sandwich ELISA is well suited to titrate GFP in solution using this antibody in combination with monoclonal anti-GFP using either form of the antibody as the capture or detection antibody. However, use the monoclonal form only for the detection of wild type or recombinant GFP as this form does not sufficiently detect 'enhanced' GFP. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin conjugated HRP. Fluorochrome conjugated polyclonal anti-GFP can be used to detect GFP by immunofluorescence microscopy in prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and can detect GFP containing inserts. Significant amplification of signal is achieved using fluorochrome conjugated polyclonal anti-GFP relative to the fluorescence of GFP alone. For immunoblotting use either alkaline phosphatase or peroxidase conjugated polyclonal anti-GFP to detect GFP or GFP containing proteins on western blots. Optimal titers for applications should be determined by the researcher.

NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).

Reviewed Applications

Read 2 reviews rated 5 using NB600-313 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% Gentamicin Sulfate

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 [HRP]

Certificate of Analysis

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

Product Specific Notices for GFP Antibody [HRP]

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 [HRP]

Customer Reviews for GFP Antibody [HRP] (2)

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Customer Images

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  • GFP Antibody [HRP]
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T cell lysate
    Species: Human
    Verified Customer | Posted 05/01/2019
    This GFP antibody works well with mGFP tagged over-expression. Because it is conjugated with HRP, secondary antibody is not needed, saves lots of time.
    GFP Antibody [HRP] NB600-313
  • Name: Gummuluru
    Application: Western Blot
    Sample Tested: HEK293T sups, Sample Amount: 100ng
    Species: Human
    Verified Customer | Posted 01/20/2012

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Protocols

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FAQs for GFP Antibody [HRP]

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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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