Product nameAnti-GFP antibody (Sepharose)
See all GFP primary antibodies
DescriptionRabbit polyclonal to GFP (Sepharose)
SpecificityThis product is a suspension of affinity purified anti-GFP antibody (ab6556) covalently linked to sepharose beads. It is reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP, CFP, RFP, and EGFP. The unit sold contains 25µg of affinity purified rabbit anti-GFP IgG cross-linked to 125 µl sepharose beads in a total volume of 250 µl buffer. The product is supplied as a 50% slurry to facilitate pipetting. Pipetting the slurry is facilitated when pipet tips are blunted by cutting 3-5mm off from the tip.
Tested applicationsSuitable for: IPmore details
Species reactivityReacts with: Species independent
Recombinant full length protein corresponding to GFP.
Database link: P42212
General notesThis product should be kept refrigerated at all times whilst in short term storage. Using sterilised equipment will reduce the risk of bacterial contamination.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Storage bufferPreservative: 0.05% Sodium azide
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThis antibody is an affinity purified rabbit anti-GFP antibody purified on an affinity chromatography column made with highly purified recombinant GFP.
Our Abpromise guarantee covers the use of ab69314 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.
We rountinely recommend using 20 µL of slurry per IP, which corresponds to 2 µg of IgG cross-linked to beads. this results in a clearly visible 10 µL bead pellet upon centrifugation. Unconjugated version: ab6556. FITC conjugated version: ab66180. Biotin conjugated version: ab69313.
RelevanceFunction: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.
Subunit structure: Monomer.
Tissue specificity: Photocytes.
Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.
Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.
Sequence similarities: Belongs to the GFP family.
Biophysicochemical properties: Absorption: Abs(max)=395 nm
Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.
- GFP antibody
- Green fluorescent protein antibody
Lane 1 & 3: IP from COS 7 cells transfected with EGFP.N-Ras (48 kDa)
Lane 2 & 4: IP from untransfected COS 7 cells
Lane 1 & 2: IP using 15 ul of rabbit anti-GFP conjugated to sepharose beads (0.5 mg IgG per ml of beads)
Lane 3 & 4: IP using 15 ul of goat anti-GFP conjugated to sepharose beads (1 mg IgG per ml of beads)
Immunoprecipitation: COS 7 cell lysates containing 100 ug of total protein in 200 ul of 0.1% SDS-RIPA buffer with addition of complete protease inhibitor were used for each immunoprecipitation. Cell lysates were incubated with anti-GFP sepharose beads for 2 hours at 4oC with rocking. Beads were washed. Proteins were eluted with 1% SDS 50 mM HEPES (pH 7.4) at 80oC (15 min). Half of the IP sample was loaded on each lane of 10 % SDS PAGE gel and gel was processed for Western blotting/ECL.
Western blot: Primary antibody: monoclonal mouse anti-GFP (ab291) at 0.2 ug/ml in 5% non-fat milk/TBS-T; 1 hour incubation at room
This product has been referenced in:
- Hsu MC et al. Protein arginine methyltransferase 3-induced metabolic reprogramming is a vulnerable target of pancreatic cancer. J Hematol Oncol 12:79 (2019). Read more (PubMed: 31324208) »
- Mishra PK et al. Cell cycle-dependent association of polo kinase Cdc5 with CENP-A contributes to faithful chromosome segregation in budding yeast. Mol Biol Cell 30:1020-1036 (2019). Read more (PubMed: 30726152) »