Overview

  • Product name
    Anti-RFP antibody (Biotin)
    See all RFP primary antibodies
  • Description
    Rabbit polyclonal to RFP (Biotin)
  • Host species
    Rabbit
  • Conjugation
    Biotin
  • Specificity
    ab34771 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 applications
    Suitable for: ELISA, WB, ICC, IHC-P, IHC-Fr, IHC-FrFlmore details
  • Immunogen

    Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.

  • General notes

    Biotin/Protein Ratio: 10-20 BAC molecules per IgG molecule Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC)

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 1% BSA, 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    This 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.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
ELISA 1/10000 - 1/80000.
WB 1/2000 - 1/10000. Detects a band of approximately 27 kDa.
ICC 1/1600.
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-Fr 1/400.
IHC-FrFl Use at an assay dependent concentration. PubMed: 24051358

Target

  • Relevance
    Fluorescent 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.
  • Alternative names
    • DsRed antibody
    • GFP like chromoprotein antibody
    • red fluorescent protein antibody
    • RFP antibody

Images

  • All lanes : Anti-RFP antibody (Biotin) (ab34771) at 1/2500 dilution

    Lane 1 : 10 µg of HeLa cell
    extract containing GFP
    recombinant protein
    Lane 2 : 10 µg
    of a HeLa lysate containing RFP recombinant protein.


    Secondary
    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.

    See Abreview

  • 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.

    See Abreview

  • Anti-RFP antibody (Biotin) (ab34771) at 1/1000 dilution
    Secondary
    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.

    See Abreview

  • 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.

    See Abreview

References

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) »
See all 46 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (neuronal)
Specification
neuronal
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton-x 100
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse epidermis and dermis)
Specification
Mouse epidermis and dermis
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 13 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Subcutaneous tissue)
Specification
Subcutaneous tissue
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate 10 mM (pH 6.0)
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 13 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Mouse embryonic stem cells transduced w/ RFP gene.)
Specification
Mouse embryonic stem cells transduced w/ RFP gene.
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 09 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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