Key Product Details

Species Reactivity

Multi-Species

Applications

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1025527
View all GFP Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant GFP
aa 2-238

Specificity

Detects GFP in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for GFP Antibody

Detection of GFP by Western Blot.

Western blot shows synthetic eGFP, recombinant RANGAP-1 tagged with eYFP, recombinant GRPuv, and crystal jelly GFP. PVDF membrane was probed with 2 µg/mL of Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42402) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for GFP at approximately 28 and recombinant RANGAP-1 tagged with eYFP at 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

GFP in HEK293 Human Cell Line Transfected with GFP.

GFP was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with GFP (green; right panel) using Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42402) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, left panel; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of GFP in HEK293 Human Cell Line Transfected with eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with protein tagged with either eGFP (filled histogram) or mCherry (open histogram) was stained with Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42402) or Mouse IgG2B Isotype Control (MAB0041, data not shown) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of GFP by Simple WesternTM.

Simple Western lane view shows eGFP and recombinant human RANGAP-1 tagged with eYFP, loaded at 0.2 mg/mL. Specific bands were detected for GFP at approximately 35 kDa and recombinant human RANGAP-1 tagged with eYFP at approximately 51 kDa (as indicated) using 10 µg/mL of Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42402). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for GFP Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line transfected with GFP

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 Human Cell Line Transfected with eGFP fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). 

Simple Western

10 µg/mL
Sample: eGFP and recombinant human RANGAP-1 tagged with eYFP

Western Blot

2 µg/mL
Sample: Synthetic eGFP, recombinant RANGAP-1 tagged with eYFP, recombinant GRPuv, and crystal jelly GFP

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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 originally isolated from the jellyfish victoria. In the presence of UV light (490-520 nm), it emits a green fluorescent color that can be used to pinpoint locations of various intracellular proteins. GFP is 238 amino acids (aa) in length. It is a globular monomer that has a tendency to dimerize. The monomer has the shape of a beta -barrel with a chromophore (aa 65-67) containing alpha -helix running up its center. +36 GFP is a superpositively charged GFP variant that can penetrate mammalian cells with potencies much greater than that of cationic peptides or modestly cationic engineered proteins.  Low molar concentrations of the protein can be observed within minutes of exposure.

Long Name

Green Fluorescent Protein

Alternate Names

eGFP, GFPuv

Additional GFP Products

Product Documents for GFP Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for GFP Antibody

For research use only

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Protocols

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