GFP antibody (Green Fluorescent Protein) Primary Antibody
This GFP antibody is un-conjugated
ELISA, Western Blotting (WB)
- For biochemical analysis of GFP-tagged fusion proteins.
- 3H9 efficiently recognizes green fluorescent proteins from Aequorea victoria such as eGFP, wtGFP, YFP, or CFP, but no red fluorescent proteins.
Fluorescent proteins (FP) are powerful tools to study protein localization and dynamics in living cells in particular the prominent green fluorescent protein (GFP) from Aequorea victoria.
Here we offer a rat monoclonal antibody (MAb) against various GFPs, designated as 3H9. The 3H9 MAb is characterized by high affinity and specificity. It is suitable for immunoblotting and ELISA.
With this versatile antibody GFP and fusions thereof become attractive tools for biochemical applications like identification of interacting proteins or Western Blotting.
- Hybridoma supernatant. The Hybridoma was cultured in protein free medium. Tissue culture supernatant was concentrated by centrifugation using a 30 kDa exclusion membrane to an antibody concentration of 1 mg/ml.
anti-Green Fluorescent Protein (GFP) antibody (Abberior STAR 635P) Primary Antibody
GFP Reactivity: Aequorea victoria FACS, ICC, IF Host: Camelid (Camelidae) Monoclonal 1H1-1B2 Abberior STAR 635P single-domain Antibody (sdAb)
- Application Notes
- Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 31 kDa (the predicted molecular weight of GFP).
IF: not working
- For Research Use only
Validation #029762 (Western Blotting)Validation ImagesFull Methods
- Moores Cancer Center, UC San Diego
- Lot Number
- Method validated
- Western Blotting
- Positive Control
- 293FT cells transduced with eGFP virus
- Negative Control
- untransduced 293FT cells
- A major band was observed at the correct molecular weight in the two positive (eGFP-transduced) cell lysates. No major bands were observed in the negative control (untransduced cells). Some additional faint bands of lower molecular weight were also observed in the positive controls, this may be because the antibody was used at 1:500 instead of the recommended 1:1000 dilution. Microscope images of transduced cells are provided to demonstrate successful eGFP-transduction of cells.
- Primary Antibody
- Antigen: Green Fluorescent Protein (GFP)
- Catalog number: ABIN398304
- Lot number: 100316
- Secondary Antibody
- Antibody: Alexa Fluor 680 Goat Anti-Rat IgG (H+L)
- Lot number: N/A
- Full Protocol
- 20 µg of total protein from prepared cell lysates was run on a 4-12% SDS-PAGE gel.
- The gel was run at 180 V for 40 minutes and transferred to a PVDF membrane using a wet-transfer apparatus.
- The membrane was rinsed in TBST for 10 min in TBST.
- The membrane was blocked in 3% BSA for 30 min.
- The membrane was incubated in 1:500 primary antibody diluted in 3% BSA at 4°C overnight.
- The membrane was washed 3 times for 10 min in TBST.
- The membrane was incubated in secondary antibody at 1:10000 for 1 hr at RT.
- The membrane was washed 3 times for 10 min in TBST.
- The membrane was imaged using a fluorescent imaging system at 679nm⁄702nm.
- Experimental Notes
- Fluorescent microscopy images are included to confirm eGFP transduction of 293FT cells.
- 1 mg/mL
- Sodium azide
- Precaution of Use
- This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Handling Advice
- Do not freeze.
- 4 °C
- Expiry Date
- 12 months
Fiskin, Bhogaraju, Herhaus, Kalayil, Hahn, Dikic: "Structural basis for the recognition and degradation of host TRIM proteins by Salmonella effector SopA." in: Nature communications, Vol. 8, pp. 14004, 2018 (PubMed).
Teixeira, Johnsen, Martínez-Montañés, Grippa, Buxó, Idrissi, Ejsing, Carvalho: "Regulation of lipid droplets by metabolically controlled Ldo isoforms." in: The Journal of cell biology, Vol. 217, Issue 1, pp. 127-138, 2018 (PubMed).
Jimenez-Orgaz, Kvainickas, Nägele, Denner, Eimer, Dengjel, Steinberg: "Control of RAB7 activity and localization through the retromer-TBC1D5 complex enables RAB7-dependent mitophagy." in: The EMBO journal, Vol. 37, Issue 2, pp. 235-254, 2018 (PubMed).
Virant, Traenkle, Maier, Kaiser, Bodenhöfer, Schmees, Vojnovic, Pisak-Lukáts, Endesfelder, Rothbauer: "A peptide tag-specific nanobody enables high-quality labeling for dSTORM imaging." in: Nature communications, Vol. 9, Issue 1, pp. 930, 2018 (PubMed).
Malik, Macartney, Hornberger, Anderson, Tovell, Prescott, Alessi: "Mechanism of activation of SGK3 by growth factors via the Class 1 and Class 3 PI3Ks." in: The Biochemical journal, Vol. 475, Issue 1, pp. 117-135, 2018 (PubMed).
Thillaiappan, Chavda, Tovey, Prole, Taylor: "Ca2+ signals initiate at immobile IP3 receptors adjacent to ER-plasma membrane junctions." in: Nature communications, Vol. 8, Issue 1, pp. 1505, 2018 (PubMed).
Duheron, Nilles, Pecenko, Martinelli, Fahrenkrog: "Localisation of Nup153 and SENP1 to nuclear pore complexes is required for 53BP1-mediated DNA double-strand break repair." in: Journal of cell science, Vol. 130, Issue 14, pp. 2306-2316, 2018 (PubMed).
Robeson, Lindblom, Wojton, Kornbluth, Matsuura: "Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators." in: The EMBO journal, Vol. 37, Issue 14, 2018 (PubMed).
Hiraga, Ly, Garzón, Hořejší, Ohkubo, Endo, Obuse, Boulton, Lamond, Donaldson: "Human RIF1 and protein phosphatase 1 stimulate DNA replication origin licensing but suppress origin activation." in: EMBO reports, Vol. 18, Issue 3, pp. 403-419, 2017 (PubMed).
Zhang, Rausch, Hastert, Boneva, Filatova, Patil, Nuber, Gao, Zhao, Cardoso: "Methyl-CpG binding domain protein 1 regulates localization and activity of Tet1 in a CXXC3 domain-dependent manner." in: Nucleic acids research, Vol. 45, Issue 12, pp. 7118-7136, 2017 (PubMed).
Toulany, Maier, Iida, Rebholz, Holler, Grottke, Jüker, Wheeler, Rothbauer, Rodemann: "Akt1 and Akt3 but not Akt2 through interaction with DNA-PKcs stimulate proliferation and post-irradiation cell survival of K-RAS-mutated cancer cells." in: Cell death discovery, Vol. 3, pp. 17072, 2017 (PubMed).
Smets, Link, Wolf, Schneider, Solis, Ryan, Meilinger, Qin, Leonhardt: "DNMT1 mutations found in HSANIE patients affect interaction with UHRF1 and neuronal differentiation." in: Human molecular genetics, Vol. 26, Issue 8, pp. 1522-1534, 2017 (PubMed).
Huber, Theiler, de Quervain, Wiens, Karangenc, Heussler, Dobbelaere, Woods: "The Microtubule-Stabilizing Protein CLASP1 Associates with the Theileria annulata Schizont Surface via Its Kinetochore-Binding Domain." in: mSphere, Vol. 2, Issue 4, 2017 (PubMed).
Kvainickas, Orgaz, Nägele, Diedrich, Heesom, Dengjel, Cullen, Steinberg: "Retromer- and WASH-dependent sorting of nutrient transporters requires a multivalent interaction network with ANKRD50." in: Journal of cell science, Vol. 130, Issue 2, pp. 382-395, 2017 (PubMed).
Morettin, Paris, Bouzid, Baldwin, Falls, Bell, Côté: "Tudor Domain Containing Protein 3 Promotes Tumorigenesis and Invasive Capacity of Breast Cancer Cells." in: Scientific reports, Vol. 7, Issue 1, pp. 5153, 2017 (PubMed).
Tawo, Pokrzywa, Kevei, Akyuz, Balaji, Adrian, Höhfeld, Hoppe: "The Ubiquitin Ligase CHIP Integrates Proteostasis and Aging by Regulation of Insulin Receptor Turnover." in: Cell, Vol. 169, Issue 3, pp. 470-482.e13, 2017 (PubMed).
Wong, Power, Miles, Tropepe: "Mutual antagonism of the paired-type homeobox genes, vsx2 and dmbx1, regulates retinal progenitor cell cycle exit upstream of ccnd1 expression." in: Developmental biology, Vol. 402, Issue 2, pp. 216-28, 2015 (PubMed).
Heinick, Husser, Himmler, Kirchhefer, Nunes, Schulte, Seidl, Rolfes, Dedman, Kaetzel, Gerke, Schmitz, Müller: "Annexin A4 is a novel direct regulator of adenylyl cyclase type 5." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2015 (PubMed).
Uhlenbrock, van Andel, Andresen, Skov: "A conserved WW domain-like motif regulates invariant chain-dependent cell-surface transport of the NKG2D ligand ULBP2." in: Molecular immunology, Vol. 66, Issue 2, pp. 418-27, 2015 (PubMed).
Groll, Emele, Poetz, Rothbauer: "Towards multiplexed protein-protein interaction analysis using protein tag-specific nanobodies." in: Journal of proteomics, 2015 (PubMed).
- Fiskin, Bhogaraju, Herhaus, Kalayil, Hahn, Dikic: "Structural basis for the recognition and degradation of host TRIM proteins by Salmonella effector SopA." in: Nature communications, Vol. 8, pp. 14004, 2018 (PubMed).
- green fluorescent protein, gfp
- The first green fluorescent protein (GFP) was isolated from Aequorea victoria and has a molecular weight of 27 kDa. Genetic mutations of GFP resulted in fluorescent proteins with different excitation and emission spectra e.g. eGFP, YFP, CFP. Fluorescent proteins became attractive tools for a variety of applications.