GFP Antibody

  
  • Specificity
    Detects GFP in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant GFP
    Ser2-Lys238
    Accession # AAB65663
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    2.5 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant GFP, see our available Western blot detection antibodies
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunocytochemistry
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 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; 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.
Intracellular Staining by Flow Cytometry
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 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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; Enhanced Green Fluorescent Protein; GFP; GFPuv; green fluorescent protein (gfp); Green Fluorescent Protein
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Goat HRP-DAB Cell & Tissue Staining Kit

CTS008 11
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ExactaChIP Chromatin Immunoprecipitation Kits
Description Application Cat# Citations Images  

ExactaChIP Chromatin IP Buffer Panel

ChIP ECP001 2
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Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 138  
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Molecular Weight Markers
Description Application Cat# Citations Images  

Full-Range Molecular Weight Marker, Biotinylated

MW001
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Secondary Antibodies
Description Application Cat# Citations Images  

Goat IgG Horseradish Peroxidase-conjugated Antibody

WB, Simple Western HAF109 13  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF017 11  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 6
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 2  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 4  
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 3  
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 4
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Donkey Anti-Goat IgG Antibody

WB AF109 5
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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