GFP Antibody

  
  • Specificity
    Detects recombinant GFPuv and eGFP in Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 454505
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant GFPuv
    Ser2-Lys238
    Accession # P42212
  • 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
    0.5 µg/mL
    See below
  • Immunoprecipitation
    2 µg/100 µg cell lysate
    293T human embryonic kidney cell line transfected with eGFP
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 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 eGFP by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line either mock transfected or transfected with eGFP. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-GFPuv/eGFP Monoclonal Antibody (Catalog # MAB42401) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for eGFP at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Immunocytochemistry GFP was detected in 2% paraformaldehyde immersion fixed HEK293 human embryonic kidney cell line transfected with GFP-expressing protein using Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42401) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007). Lower panel shows transfected cells by GFPuv auto-fluorescence (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
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 and 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).To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
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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  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Primary Antibodies
Description Application Cat# Citations Images  

GFP Antibody

WB, Flow, ICC MAB42401R  
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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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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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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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™ NL493-conjugated Antibody

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

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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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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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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