GFP Antibody

Catalog # Availability Size / Price Qty
MAB42401
MAB42401-SP
GFP Antibody in Data
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GFP Antibody Summary

Specificity
Stains eGFP transfectants but not irrelevant transfectants in flow cytometry. Detects recombinant GFPuv in Western blots. For optimal Western blot results, we recommend R&D Systems Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) or Novus Biologicals Mouse Anti-GFP Monoclonal Antibody (Catalog # NBP2-2211).
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
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

Immunocytochemistry View Larger

GFP in HEK293 Human Cell Line. 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. View Larger

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 IgG1Isotype 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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

FAQs

No product specific FAQs exist for this product, however you may

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Cell and Tissue Staining Kits

Isotype Controls

Primary Antibodies

Reconstitution Buffers

Secondary Antibodies

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