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).

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for GFP Antibody

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.

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.

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

8-25 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line transfected with eGFP

Immunoprecipitation

2 µg/100 µg cell lysate
Sample: 293T human embryonic kidney cell line transfected with eGFP

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 human embryonic kidney cell line transfected with eGFP was fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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.

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

Product Datasheets

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COA

SDS

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Product Specific Notices for GFP Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

GFP Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB42401

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

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Product Specifications

Immunogen

Specificity