| |
code |
price |
size |
| |
5f8 |
€170.00 |
100 µg |
| Description |
Rat monoclonal [5F8] to monomeric 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.
Use this rat monoclonal antibody (MAb) against monomeric RFPs, designated as RFP antibody [5F8]. The 5F8 MAb is characterized by high affinity and specificity against the monomeric 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 RFP 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 RFp 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. |
| |
