详细说明
Specificity
Detects recombinant GFPuv and eGFP in Western blots.
Source
Recombinant Monoclonal Mouse IgG 1 Clone # 454505R
Purification
Protein A or G purified from cell culture supernatant
Immunogen
E.coli-derived recombinant GFPuv
Ser2-Lys238
Accession # P42212Formulation
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
ConcentrationSample
Western Blot
2 µg/mL
See below
Flow Cytometry
0.25 µg/10 6 cells
See below
Immunocytochemistry
5-25 µg/mL
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | 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 # ). A specific band was detected for GFP at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
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 either (A) Mouse Anti-GFP Monoclonal Antibody (Catalog # MAB42401) or (B) Mouse IgG1 Isotype Control (Catalog # ) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # ). |
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 # ) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in GFP-positive cells. View our protocol for . |
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
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