|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 # HAF018). A specific band was detected for GFP at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|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 # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).|
|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 # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in GFP-positive cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
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.
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.