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Products > Monoclonal antibodies > RFP antibody

  code price size
  5f8 €170.00 100 µg
Description Rat monoclonal [5F8] to Multiple Red Fluorescent Proteins.
For biochemical and cell biological analysis of RFP-tagged fusion proteins.
Product Type Primary antibody
Specificity 5F8 recognizes various Red fluorescent proteins derived from DsRed and RFP-tagged fusion proteins.
Isotype IgG2a
Relevance 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.
Tested Applications and Recommended Dilutions

Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight RFP: 30 kDa).

primary antibody: 0.2 µg 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

IP, Co-IP

For immunoprecipitation 4µg 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)  

Protocol (PDF)

Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Reference Rottach, A et al. Generation and characterization of a rat monoclonal antibody specific for multiple red fluorescent proteins. Hybridoma (2008). PubMed: 18788935
Form 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.
Storage Buffer Preservative: 0.01% Sodium Azide
Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
Concentration 1 mg/ml
Storage instructions Shipped at RT. Upon delivery aliquot and store at -20°C. Store at 4°C short term (1-2 weeks). Avoid freeze / thaw cycles.
 

Only for research applications, not for diagnostic or therapeutic use.