Key Product Details

Species Reactivity

Non-species specific

Applications

Validated:

Knockout Validated, Western Blot, ELISA, Fluorophore-linked immunosorbent assay, Immunocytochemistry/ Immunofluorescence, Dot Blot, Knockdown Validated

Cited:

Immunocytochemistry/ Immunofluorescence

Label

DyLight 488 (Excitation = 493 nm, Emission = 518 nm)

Antibody Source

Polyclonal Goat IgG
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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 or Rat serum proteins.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

Store vial at 4C prior to restoration. 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 temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

GFP Dylight(TM) 488 Conjugated 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 and purified and partially purified Green Fluorescent Protein (Aequorea victoria)

Scientific Data Images for GFP Antibody [DyLight 488]

Western Blot Detection of GFP Using DyLight 488 Conjugated Antibody

Western Blot Detection of GFP Using DyLight 488 Conjugated Antibody

Lane 1: His-Sumo-GFP. Lane: Molecular Weight. Lane 2: Beta-Galactosidase (negative control). Lane 3: recombinant GFP control protein. Load: 35 ug per lane. Primary antibody: none. Secondary antibody: DyLight (TM) 488 conjugated anti-GFP goat secondary antibody at 1:5,000. Block for2 hr at RT. Predicted/Observed size: 27kDa/54kDa, 27kDa for rGFP/~45kDa His-Sumo-GFP.
Comparison of Fluorescence Absorption and Emission Spectra for DyLight 488 and Alexa Fluor 488 Conjugates

Comparison of Fluorescence Absorption and Emission Spectra for DyLight 488 and Alexa Fluor 488 Conjugates

Comparison of fluorescence absorption and emission spectra for DyLight (TM) 488 and Alexa Fluor 488 in PBS, pH7.2. The emission spectra for this DyLight (TM) conjugate match the principle output wavelengths of most common fluorescence instrumentation.
Immunofluorescent Staining of Human Breast Carcinoma Tissue Using DyLight 488 Conjugated GFP Antibody

Immunofluorescent Staining of Human Breast Carcinoma Tissue Using DyLight 488 Conjugated GFP Antibody

Tissue: human breast carcinoma. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: Anti-Histone and Anti-Tubulin antibody at 10 ug/mL for 1 h at RT. Secondary antibody: DyLight (R) 488 conjugate and DyLight (R) 549 conjugate goat secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone is nuclear and Tubulin is cytoplasmic. Staining: Anti-Histone detection using a DyLight (R) 488 conjugate (green) fluorescent signal and Anti-Tubulin was detected using a DyLight (R) 549 conjugate (red) fluorescent signal. Nuclei were counter-stained using DAPI (blue).
Use of DyLight 488 Conjugated GFP Antibody in Western Blot

Use of DyLight 488 Conjugated GFP Antibody in Western Blot

Western Bot of Goat Anti-GFP (GOAT) Antibody DyLight(TM) 488 Conjugated. Lane 1: Opal Prestained Molecular Weight Marker

Applications for GFP Antibody [DyLight 488]

Application
Recommended Usage

ELISA

1:100 - 1:2000

Fluorophore-linked immunosorbent assay

1:100 - 1:2000

Immunocytochemistry/ Immunofluorescence

>1:5000

Western Blot

1:10000
Application Notes
This product has been tested by dot blot and western blot. The emission spectra for this DyLight(TM) conjugate match the principle output wavelengths of most common fluorescence instrumentation.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Reconstitution

Reconstitute with 100 ul deionized water (or equivalent)

Formulation

Lyophilized from 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

LYOPH mg/ml

Shipping

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

Stability & Storage

Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 [DyLight 488]

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 [DyLight 488]



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

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 [DyLight 488]

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Protocols

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

FAQs for GFP Antibody [DyLight 488]

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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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