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Key Product Details

Species Reactivity

Non-species specific

Applications

Validated:

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

Cited:

ELISA

Label

Alkaline Phosphatase

Antibody Source

Polyclonal Goat IgG

Concentration

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

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

Store vial at 4C before opening. DO NOT FREEZE. This product is stable at 4C as an undiluted liquid. Dilute only prior to immediate use. Freezing alkaline phosphatase conjugates will result in a substantial loss of enzymatic activity.

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-Alkaline Phosphatase and purified and partially purified Green Fluorescent Protein (Aequorea victoria) Serum

Scientific Data Images for GFP Antibody [Alkaline Phosphatase]

Western Blot Detection of GFP Using Alkaline Phosphatase Conjugated Antibody

Western Blot Detection of GFP Using Alkaline Phosphatase Conjugated Antibody

Western Blot of GFP antibody [Alkaline Phosphatase]. Lane 1: GFP. Lane 2: none. Load: 50 ng per lane.Primary antibody: none.Secondary antibody: Alkaline Phosphatase GFP secondary antibody at 1:1000 for 60 min at RT.Block for 30 min RT.Predicted/Observed size: 28 kDa for GFP. Other band(s): none.
Dot Blot Analysis Using Alkaline Phosphatase Conjugated GFP Antibody

Dot Blot Analysis Using Alkaline Phosphatase Conjugated GFP Antibody

Dot Blot of GFP Antibody Alkaline Phosphatase Conjugated. Antigen: GFP. Load: Lane 1 - 100ng Lane 2 - 33.33ng Lane 3 - 11.11ng Lane 4 - 3.70ng Lane 5 - 1.23ng. Primary antibody: n/a. Secondary antibody: GFP Antibody [Alkaline Phosphatase]at 1:1,000 for 60 min at RT. Block for 60 min at RT. Visualized using Alkaline Phosphatase Substrate.

Applications for GFP Antibody [Alkaline Phosphatase]

Application
Recommended Usage

ELISA

1:2000-1:8000

Immunohistochemistry

1:200-1:1000

Immunohistochemistry-Paraffin

1:10-1:500

Simple Western

1:10

Western Blot

1:500-1:2500
Application Notes
This product is designed to detect GFP and its variants. This product Alkaline Phosphatase conjugated antibody has been tested by ELISA, dot blot, and western blot. This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP. 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 antibodies. 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

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.05 M Tris Chloride, 0.15M Sodium Chloride, 0.001M Magnesium Chloride, 0.0001M Zinc Chloride, 50% (v/v) Glycerol; pH 8.0, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative

0.1% 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. Do not freeze.

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 [Alkaline Phosphatase]

Certificate of Analysis

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

Product Specific Notices for GFP Antibody [Alkaline Phosphatase]

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.

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