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GFP Antibody

R&D Systems, part of Bio-Techne | Catalog # AF4240

R&D Systems, part of Bio-Techne
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AF4240
AF4240-SP

Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse, Rat

Applications

Validated:

CyTOF-ready, Immunocytochemistry, Immunoprecipitation, Intracellular Staining by Flow Cytometry, Simple Western, Western Blot

Cited:

Bioassay, Co-IP, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant GFPuv
Ser2-Lys238
Accession # P42212

Specificity

Detects GFP in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for GFP Antibody

Detection of GFP antibody by Western Blot.

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.
GFP antibody in HEK293 human embryonic kidney cells transfected with GFP by Immunocytochemistry (ICC).

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 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, middle panel; 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.
Detection of GFP antibody in HEK293 human embryonic kidney cells transfected with GFP.

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).

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

1-15 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cells transfected with wild type GFP

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant GFP, see our available Western blot detection antibodies

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Immersion fixed HEK293 human embryonic kidney cells transfected with wild type GFP

Simple Western

2.5 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with eGFP-tagged proteins

Western Blot

1 µg/mL
Sample: NS0 mouse myeloma cell line either transfected with eGFP-tagged proteins
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 4 reviews rated 4.3 using AF4240 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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

Additional GFP Products

Product Documents for GFP Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for GFP Antibody

For research use only

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