Product nameAnti-RFP antibody (Biotin)
See all RFP primary antibodies
DescriptionRabbit polyclonal to RFP (Biotin)
Specificityab34771 recognises RFP. The antibody is predicted to react with all RFP variants from Discosoma (including DsRed, TdTomato, mOrange etc.) based on sequence homology and polyclonality of the antibody. However, such crossreactivity has not been QC tested.
Tested applicationsSuitable for: ELISA, WB, ICC, IHC-P, IHC-Fr, IHC-FrFlmore details
Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.
Biotin/Protein Ratio: 10-20 BAC molecules per IgG molecule Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 1% BSA, 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThis product was prepared from monospecific antiserum by immunoaffinity chromatography using Red Fluorescent Protein (Discosoma) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. No reaction was observed against Human, Mouse or Rat serum proteins. ELISA was used to confirm specificity at less than 0.1% of target signal.
Our Abpromise guarantee covers the use of ab34771 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||1/10000 - 1/80000.|
|WB||1/2000 - 1/10000. Detects a band of approximately 27 kDa.|
|IHC-P||1/1000 - 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Endogenous tissue biotin should be considered as a source of false-positive staining when immunohistochemical or histochemical techniques which use avidin or streptavidin reagents or anti-biotin antibodies as components of the detection system, are applied to tissue sections. We recommned blocking the endogenous biotins.
|IHC-FrFl||Use at an assay dependent concentration. PubMed: 24051358|
RelevanceFluorescent proteins have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions.
- DsRed antibody
- GFP like chromoprotein antibody
- red fluorescent protein antibody
- RFP antibody
All lanes : Anti-RFP antibody (Biotin) (ab34771) at 1/2500 dilution
Lane 1 : 10 µg of HeLa cell
extract containing GFP
Lane 2 : 10 µg
of a HeLa lysate containing RFP recombinant protein.
All lanes : IRDye™800 conjugated Goat anti-Rabbit IgG at 1/5000 dilution
Observed band size: 27 kDa why is the actual band size different from the predicted?
IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
ab34771 staining mouse subcutaneous tissue that had been injected with a mixture of cells including mouse embryonic stem cells previously transduced with a retrovirus that expresses RFP.
The sample was fixed with paraformaldehyde and a heat mediated antigen retreival step using Sodium citrate 10 mM (pH 6.0) was performed. 1.5% serum was used as the blocking agent for 30 minutes at 20°C. The primary antibody was diluted 1/400 in 1xPBS with 1.5% normal serum and incubated for 1 hour at 20°C. A Biotinylated goat anti-rabbit IgG (H+L) antibody was used as the secondary.
ab34771 staining RFP in transfected Mouse N2a cells by ICC (Immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 5% serum for 2 hours at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 2% BSA) for 12 hours at 4°C. An Alexa Fluor®546-Streptavidin -conjugated anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Anti-RFP antibody (Biotin) (ab34771) at 1/1000 dilution
Peroxidase streptavidin secondary antibody at 1/40000 dilution
ab34771 staining mouse epidermis that had been damaged and injected with a mixture of cells inlcuding mouse embryonic stem cells previously transduced with a retrovirus that expresses RFP. The color is developed with DAB.
The sample was acetone fixed and incubated with ab34771 diluted 1/400 for 1 hour at 20°C. A Biotinylated goat anti-rabbit IgG (H+L) antibody was used as the secondary.
ICC image of ab34771 staining murine embryonic stem cells, transduced with RFP gene, cultured over mouse embryonic fibroblast. The cells were PFA fixed and blocked with 1.5% serum for 45 minutes at 20°C. The primary antibody was diluted 1/1600 and incubated for 1 hour and 15 minutes at 20°C. A biotinylated goat anti-rabbit IgG (H+L) was used as the secondary.
This product has been referenced in:
- Kothari P et al. Contractility kits promote assembly of the mechanoresponsive cytoskeletal network. J Cell Sci 132:N/A (2019). Read more (PubMed: 30559246) »
- Hashimoto-Torii K et al. Detection of local and remote cellular damage caused by spinal cord and peripheral nerve injury using a heat shock signaling reporter system. IBRO Rep 5:91-98 (2018). Read more (PubMed: 30480161) »