GFP Antibody

Recombinant Monoclonal Antibody.
  
  • Specificity
    Detects recombinant GFPuv and eGFP in Western blots.
  • Source
    Recombinant Monoclonal Mouse IgG1 Clone # 454505R
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E.coli-derived recombinant GFPuv
    Ser2-Lys238
    Accession # P42212
  • Formulation
    Supplied as a solution in PBS. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunocytochemistry
    5-25 µg/mL
    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 Human GFP by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with eGFP. PVDF membrane was probed with 2 µg/mL of Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42401R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for GFP at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of eGFP in HEK293 Human Cell Line Transfected with eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with eGFP was stained with either (A) Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42401) or (B) Mouse IgG1 Isotype Control (Catalog # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).
Immunocytochemistry
GFP in HEK293 Human Cell Line. GFP was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with GFP (green, upper panel) using Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42401R) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, lower panel; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in GFP-positive cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. 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 opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
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-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 127  
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Mouse IgG1 Isotype Control

Ctrl MAB002R
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 25  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse IgG HRP-conjugated Antibody

WB HAF018 5  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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