anti-GFP antibody (Green Fluorescent Protein)

Details for Product anti-GFP Antibody No. ABIN398304
Independently validated
Aequorea victoria
(481), (10), (6), (5), (4), (2), (2), (2), (1), (1), (1)
(200), (158), (80), (41), (13), (7), (2)
Clonality (Clone)
Monoclonal ()
This GFP antibody is un-conjugated
(29), (29), (21), (15), (13), (3), (3), (3), (3), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
ELISA, Western Blotting (WB)
(399), (233), (134), (118), (99), (40), (40), (38), (30), (25), (13), (12), (10), (9), (7), (7), (4), (3), (3), (1), (1), (1), (1), (1)
Pubmed 21 references available
Quantity 100 μL
Shipping to United States ( )
Availability Will be delivered in 4 to 5 Business Days
'Independent Validation' Badge
Lot Number 100316
Method validated Western Blotting
Positive Control 293FT cells transduced with eGFP virus
Negative Control untransduced 293FT cells
Notes 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.
Purpose For biochemical analysis of GFP-tagged fusion proteins.
Clone 3H9
Isotype IgG2a
Specificity 3H9 efficiently recognizes green fluorescent proteins from Aequorea victoria such as eGFP, wtGFP, YFP, or CFP, but no red fluorescent proteins.
Characteristics 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.
Purification 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.
Background 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.
Research Area Tags/Labels
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

Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Preservative 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.
Storage 4 °C
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for anti-GFP antibody (Green Fluorescent Protein) (ABIN398304) Western Blot analysis using anti-GFP antibody 3H9. Primary antibody: 0.2 ug of GFP an...
Product cited in: Hamacher-Brady, Brady: "Bax/Bak-Dependent, Drp1-Independent Targeting of XIAP into Inner-Mitochondrial Compartments Counteracts Smac-Dependent Effector Caspase Activation." in: The Journal of biological chemistry, 2015 (PubMed).

Qin, Wolf, Liu et al.: "DNA methylation requires a DNMT1 ubiquitin interacting motif (UIM) and histone ubiquitination." in: Cell research, Vol. 25, Issue 8, pp. 911-29, 2015 (PubMed).

Heinick, Husser, Himmler et al.: "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 et al.: "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 et al.: "Towards multiplexed protein-protein interaction analysis using protein tag-specific nanobodies." in: Journal of proteomics, 2015 (PubMed).

Reiner, Hoefle, Hückelhoven: "A barley SKP1-like protein controls abundance of the susceptibility factor RACB and influences the interaction of barley with the barley powdery mildew fungus." in: Molecular plant pathology, 2015 (PubMed).

Wong, Power, Miles et al.: "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).

Jokinen, Lahtinen, Marttinen et al.: "Quantitative changes in Gimap3 and Gimap5 expression modify mitochondrial DNA segregation in mice." in: Genetics, Vol. 200, Issue 1, pp. 221-35, 2015 (PubMed).

Chen, Tibroni, Sauter et al.: "Modest attenuation of HIV-1 Vpu alleles derived from elite controller plasma." in: PLoS ONE, Vol. 10, Issue 3, pp. e0120434, 2015 (PubMed).

Olazabal-Herrero, García-Santisteban, Rodríguez: "Structure-function analysis of USP1: insights into the role of Ser313 phosphorylation site and the effect of cancer-associated mutations on autocleavage." in: Molecular cancer, Vol. 14, Issue 1, pp. 33, 2015 (PubMed).

Baldwin, Bejide, Trinkle-Mulcahy et al.: "Identification of the PRMT1v1 and PRMT1v2 specific interactomes by quantitative mass spectrometry in breast cancer cells." in: Proteomics, Vol. 15, Issue 13, pp. 2187-97, 2015 (PubMed).

Scarfone, Venturetti, Hotz et al.: "Asymmetry of the budding yeast Tem1 GTPase at spindle poles is required for spindle positioning but not for mitotic exit." in: PLoS genetics, Vol. 11, Issue 2, pp. e1004938, 2015 (PubMed).

Traenkle, Emele, Anton et al.: "Monitoring interactions and dynamics of endogenous beta-catenin with intracellular nanobodies in living cells." in: Molecular & cellular proteomics : MCP, Vol. 14, Issue 3, pp. 707-23, 2015 (PubMed).

Poças, Branco-Santos, Herrera et al.: "α-Synuclein modifies mutant huntingtin aggregation and neurotoxicity in Drosophila." in: Human molecular genetics, Vol. 24, Issue 7, pp. 1898-907, 2015 (PubMed).

Galili, Dylla, Lüdke et al.: "Converging circuits mediate temperature and shock aversive olfactory conditioning in Drosophila." in: Current biology : CB, Vol. 24, Issue 15, pp. 1712-22, 2014 (PubMed).

Anton, Bauer, Keck et al.: "A p38 substrate-specific MK2-EGFP translocation assay for identification and validation of new p38 inhibitors in living cells: a comprising alternative for acquisition of cellular p38 inhibition data." in: PLoS ONE, Vol. 9, Issue 4, pp. e95641, 2014 (PubMed).

Kríz, Pospíchalová, Masek et al.: "?-arrestin promotes Wnt-induced low density lipoprotein receptor-related protein 6 (Lrp6) phosphorylation via increased membrane recruitment of Amer1 protein." in: The Journal of biological chemistry, Vol. 289, Issue 2, pp. 1128-41, 2014 (PubMed).

Lestini, Laptenok, Kühn et al.: "Intracellular dynamics of archaeal FANCM homologue Hef in response to halted DNA replication." in: Nucleic acids research, Vol. 41, Issue 22, pp. 10358-70, 2013 (PubMed).

Becker, Allmann, Hofstätter et al.: "Direct homo- and hetero-interactions of MeCP2 and MBD2." in: PLoS ONE, Vol. 8, Issue 1, pp. e53730, 2013 (PubMed).

Lee, Fischer: "Drosophila Tel2 is expressed as a translational fusion with EpsinR and is a regulator of wingless signaling." in: PLoS ONE, Vol. 7, Issue 9, pp. e46357, 2012 (PubMed).

Garcia-Santisteban, Zorroza, Rodriguez: "Two nuclear localization signals in USP1 mediate nuclear import of the USP1/UAF1 complex." in: PLoS ONE, Vol. 7, Issue 6, pp. e38570, 2012 (PubMed).

Primary Antibody
  • Antigen: Green Fluorescent Protein (GFP)
  • Catalog number: ABIN398304
  • Supplier: Chromotek
  • Supplier catalog number: 3H9
  • Lot number: 100316
Secondary Antibody
  • Antibody: Alexa Fluor 680 Goat Anti-Rat IgG (H+L)
  • Supplier: Life Technologies
  • Catalog number: A-21096
  • Lot number: N/A
  • Positive control: 293FT cells transduced with eGFP virus
  • Negative control: untransduced 293FT cells
  • 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.
Validation Images
Western Blotting validation image for anti-Green Fluorescent Protein (GFP) antibody (ABIN398304) Figure 1: A. Western blot of lysates from untransduced 293FT cells (Lane 2) and 293FT...
Catalog No. ABIN398304
$ 254.38
Plus shipping costs $45.00
100 μL
$ 254.38

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