Red Fluorescent Protein (RFP) antibody

Antigen: Red Fluorescent Protein (RFP)

Clonality: Monoclonal (5F8)

Host: Rat

Application: Western Blotting (WB), Immunoprecipitation (IP), Immunofluorescence (IF)

Catalog no.: ABIN334653

Additional Information

Format: Liquid, Supernatant

Isotype: IgG2a  (Matching secondary antibodies)

Clone: 5F8

Description: Fluorescent proteins (FP) are powerful tools to study protein localization and dynamics in living cells In addition to the prominent green fluorescent protein (GFP) from Aequorea victoria, a red fluorescent protein (DsRed) was discovered in the coral Discosoma sp. Mutagenesis of the red progenitor have resulted in several monomeric red FPs (mRFP1, mCherry, mOrange, mPlum, etc) which become attractive tools for a number of applications. For example the combination of green and red fusion proteins allows multicolor tracking as well as protein-protein interaction studies in living cells. Here we offer a rat monoclonal antibody (MAb) against multiple red FPs, designated as multi-red 5F8. The 5F8 MAb is characterized by high affinity and specificity against the DsRed derivatives and corresponding fusion proteins. It is suitable for ELISA, immunoblotting, immunoprecipitation, and immunofluorescence assays. With this versatile antibody red fluorescent proteins and fusions thereof become attractive tools for biochemical applications like identification of interacting proteins or enzymatic analyses as well as for microscopic analyses.

Specificity: 5F8 recognizes various Red fluorescent proteins derived from DsRed and RFP-tagged fusion proteins.

Application Details

Application Notes: Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight RFP: 30 kDa). Immunoprecipitation: For immunoprecipitation 4ug of multi-red antibody 5F8 used incubation for 1 h at RT. A 50 kDa band is precipitated which is IgG heavy chain whilst the 30kDa band is PCNA.Immunofluorescence: HeLa cells stably expressing mCherry-PCNA, 3,7% PFA fixed. Used at a dilution of 1/1000. Secondary antibody: donkey-anti-rat-Alexa488 (1/400)

Concentration: 1mg/ml

Buffer: Liquid, hybridoma supernatant. The hybridoma was cultured in protein free medium. Tissue culture supernatant was concentrated by centrifugation using a 30 kD exclusion membrane to an antibody concentration of 1 mg/ml. Preservative: 0.01% Sodium Azide

Storage: Shipped at 4 degree C. Upon delivery aliquot and store at -20 degree C. Store at +4 degree C short term (1-2 weeks). Avoid freeze / thaw cycles.

Research Area: Tags/Labels

Restrictions: For Research Use only

Images

primary antibody: 0.2 ug of multi-red antibody 5F8 incubation for 1 h at RT secondary antibody: anti-rat-HRP (Jackson ImmunoResearch Europe) detection with ECL Western Blot Detection Kit

Publications

Publications: Rottach, Kremmer, Nowak et al.: "Generation and characterization of a rat monoclonal antibody specific for multiple red fluorescent proteins." in: Hybridoma (2005), Vol. 27, Issue 5, pp. 337-43, 2008 (PubMed).

Bakondi, Spees: "Human CD133-derived bone marrow stromal cells establish ectopic hematopoietic microenvironments in immunodeficient mice." in: Biochemical and biophysical research communications, 2010 (PubMed).

Laxman, Sutter, Tu: "Behavior of a metabolic cycling population at the single cell level as visualized by fluorescent gene expression reporters." in: PLoS ONE, Vol. 5, Issue 9, pp. e12595, 2010 (PubMed).

Korzeniowski, Manjarra: "Activation of STIM1-Orai1 Involves an Intramolecular Switching Mechanism." in: Science signaling, Vol. 3, Issue 148, pp. ra82, 2010 (PubMed).

Qin, Leonhardt, Spada: "Usp7 and Uhrf1 control ubiquitination and stability of the maintenance DNA methyltransferase Dnmt1." in: Journal of cellular biochemistry, 2010 (PubMed).

Goldmann, Overlack, Wolfrum et al.: "PTC124 mediated translational read-through of a nonsense mutation causing Usher type 1C." in: Human gene therapy, 2011 (PubMed).

Chuhma, Tanaka, Hen et al.: "Functional connectome of the striatal medium spiny neuron." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 31, Issue 4, pp. 1183-92, 2011 (PubMed).

Speck, Arndt, Ma: "Efficient phage display of intracellularly folded proteins mediated by the TAT pathway." in: Protein engineering, design & selection : PEDS, 2011 (PubMed).

Frauer, Rottach, Meilinger et al.: "Different Binding Properties and Function of CXXC Zinc Finger Domains in Dnmt1 and Tet1." in: PLoS ONE, Vol. 6, Issue 2, pp. e16627, 2011 (PubMed).

Tags/Labels: Rancz, Franks, Schwarz et al.: "Transfection via whole-cell recording in vivo: bridging single-cell physiology, genetics and connectomics." in: Nature neuroscience, Vol. 14, Issue 4, pp. 527-32, 2011 (PubMed). Method employed by authors: Immunohistochemistry (IHC) (1:1000).

Ta?nnesen, Parish, Sa?rensen et al.: "Functional integration of grafted neural stem cell-derived dopaminergic neurons monitored by optogenetics in an in vitro Parkinson model." in: PLoS ONE, Vol. 6, Issue 3, pp. e17560, 2011 (PubMed). (1:1000). Further details: Slices were incubated 15 h

Graewe, Rankin, Lehmann et al.: "Hostile Takeover by Plasmodium: Reorganization of Parasite and Host Cell Membranes during Liver Stage Egress." in: PLoS pathogens, Vol. 7, Issue 9, pp. e1002224, 2011 (PubMed).

Uhrig, Moorhead: "Two ancient bacterial-like PPP family phosphatases from Arabidopsis are highly conserved plant proteins that possess unique properties." in: Plant physiology, Vol. 157, Issue 4, pp. 1778-92, 2011 (PubMed).

Petrovská, Cenklová, Pochylová et al.: "Plant Aurora kinases play a role in maintenance of primary meristems and control of endoreduplication." in: The New phytologist, Vol. 193, Issue 3, pp. 590-604, 2012 (PubMed).

Yoshihara, Takahashi, Nishimura et al.: "5T4 Glycoprotein Regulates the Sensory Input-Dependent Development of a Specific Subtype of Newborn Interneurons in the Mouse Olfactory Bulb." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 32, Issue 6, pp. 2217-2226, 2012 (PubMed). (1:1000).

Ingmundson, Nahar, Brinkmann et al.: "The exported Plasmodium berghei protein IBIS1 delineates membranous structures in infected red blood cells." in: Molecular microbiology, 2012 (PubMed). (1:1000).

Mu, Berti, Masserdotti et al.: "SoxC Transcription Factors Are Required for Neuronal Differentiation in Adult Hippocampal Neurogenesis." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 32, Issue 9, pp. 3067-3080, 2012 (PubMed). (1:300).