Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse, Rat

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, Immunoprecipitation, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western, Immunoprecipitation, Bioassay, Co-IP, Purification

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all GFP Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant GFPuv
Ser2-Lys238
Accession # P42212

Specificity

Detects GFP in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for GFP Antibody

Detection of GFP antibody by Western Blot.

Detection of GFP by Western Blot.

Western blot shows lysates of NS0 mouse myeloma cell line either mock transfected or transfected with eGFP-tagged EDG6. PVDF membrane was probed with 1 µg/mL of Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GFP at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

GFP antibody in HEK293 human embryonic kidney cells transfected with GFP by Immunocytochemistry (ICC).

GFP in HEK293 human embryonic kidney cells transfected with GFP.

GFP was detected in immersion fixed HEK293 human embryonic kidney cells transfected with GFP (green) using Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, middle panel; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm of GFP-positive cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of GFP antibody in HEK293 human embryonic kidney cells transfected with GFP.

Detection of GFP in HEK293 human embryonic kidney cells transfected with GFP.

HEK293 human embryonic kidney cells transfected with GFP was stained with Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody only (A, Catalog # F0108) or with Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by Secondary Antibody (B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. Quadrant markers were set based on control antibody staining (Catalog # AB-108-C).

Detection of GFP antibody by Simple WesternTM.

Detection of GFP by Simple WesternTM.

Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line either mock transfected (-) or transfected with eGFP-tagged EDG6 (+), loaded at 0.2 mg/mL. A specific band was detected for GFP at approximately 114 kDa (as indicated) using 2.5 µg/mL of Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

GFP Antibody

Simple Western: GFP Antibody [Unconjugated] [AF4240] -

Simple Western: GFP Antibody [Unconjugated] [AF4240] - Design for a membrane-localized ACE2 expression system. (A) Our ACE2 construct is driven by a CMV promoter followed by the first 25 residues of ACE2 containing the leader sequence that direct ACE2 to the plasma membrane. This is followed by a 3xHA tag linked to the remainder of ACE2 (20-805) and a C-terminal sfGFP. Both 3xHA and sfGFP fusions are separated from ACE2 by flexible 3xGGGGS linkers. (B) The ACE2 fusion protein is designed to be embedded in the plasma membrane where it can perform extracellular carboxypeptidase-mediated metabolism and its levels can be detected by cell staining with antibodies to HA. (C) Lysates from untransfected or 3xHA-ACE2-sfGFP-transfected HEK293 cells were analyzed by automated Jess capillary immunoassay using antibodies to HA, GFP, and two ACE2 antibodies. (D) Confocal fluorescence microscopy of HEK cells transfected with 3xHA-ACE2-sfGFP and stained with HA and the nuclear stain DAPI. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/37644110), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GFP by Western Blot

Detection of GFP by Western Blot

GFP-rMSCs following in vitro stress assays. (A) Trypsinized and re-plated GFP-rMSCs at 24 h following hydrogen peroxide treatment. PSF-treated cells were much more abundant and had begun dividing, but GFP-rMSCs not treated with PSF appeared to have lasting negative effects of H2O2 exposure. (B) Lactate dehydrogenase released into medium of the hydrogen peroxide-treated cells was a means to measure cell death. (C) Changes in expression of stress-related proteins Hif-1a, HSP70, and SOD1 in GFP-rMSCs following hydrogen peroxide treatment. * Indicates p < 0.5. Experiments were repeated twice. Bar =100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35054878), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for GFP Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

1-15 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cells transfected with wild type GFP

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant GFP, see our available Western blot detection antibodies

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Immersion fixed HEK293 human embryonic kidney cells transfected with wild type GFP

Simple Western

2.5 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with eGFP-tagged proteins

Western Blot

1 µg/mL
Sample: NS0 mouse myeloma cell line either transfected with eGFP-tagged proteins

Reviewed Applications

Read 4 reviews rated 4.3 using AF4240 in the following applications:

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

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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 Aequorea 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. GFPuv is the Aequorea sequence with three aa substitutions; Phe to Ser at # 99, Met to Thr at # 153, and Val to Ala at # 163. This form expresses faster and is 18-fold brighter than native GFP; excitation peaks at 395 nm and emission at 508 nm.

Long Name

Green Fluorescent Protein

Alternate Names

eGFP, GFPuv

UniProt

P42212

Additional GFP Products

Product Documents for GFP Antibody

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for GFP Antibody

For research use only

Customer Reviews for GFP Antibody (4)

4.3 out of 5
4 Customer Ratings
5 Stars
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4 Stars
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3 Stars
25%
2 Stars
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1 Stars
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • GFP Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Brain (cross-section through blood vessel)
    Species: Mouse
    Verified Customer | Posted 09/17/2018
    GFP Antibody AF4240
  • GFP Antibody
    Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: HEK293 human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 01/17/2018
    GFP Antibody AF4240
  • GFP Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Liver cells
    Species: Mouse
    Verified Customer | Posted 10/27/2017
    GFP Antibody AF4240
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 20484048
    Species: Other
    Verified Customer | Posted 01/08/2015

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Protocols

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