| 货号 | AF4240-SP |
| 别名 | eGFP; Enhanced Green Fluorescent Protein; GFPuv; green fluorescent protein (gfp); Green Fluorescent Protein | 全称 | Green Fluorescent Protein |
| 应用 | Western Blot,Simple Western,Immunoprecipitation,CyTOF-ready,Immunocytochemistry,Intracellular Staining by Flow Cytometry |
| 目标/特异性 | Detects GFP in direct ELISAs and Western blots. |
| 使用方法 | Western Blot: 1 µg/mL Simple Western: 2.5 µg/mL Immunoprecipitation: 25 µg/mL Immunocytochemistry: 5-15 µg/mL Intracellular Staining by Flow Cytometry: 0.25 µg/106cells |
| 来源 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 产品组分 |
| 供应商 | R&D Systems |
| 应用文献 | |
| R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions. Lymphatic endothelial progenitors originate from plastic myeloid cells activated by toll-like receptor-4 | |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | E. coli-derived recombinant GFP Ser2-Lys238 Accession # AAB65663 |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | Green fluorescent protein (GFP) is a 27 kDa protein originally isolated from the jellyfish Aequoreavictoria. 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. |
| 运输条件 | Blue Ice |
| 存放说明 | 4℃ |
| 参考文献 |
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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 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 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; 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 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). | |
Detection of GFP by Simple WesternTM. Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line either mock transfected (-) or transfected with eGFP-tagged EDG6 (+), loaded at 0.2 mg/mL. A specific band was detected for GFP at approximately 114 kDa (as indicated) using 2.5 µg/mL of Goat Anti-GFP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4240) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. |
