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RFP anticorps (Red Fluorescent Protein) Primary Antibody

RFP Reactivité: Discosoma ELISA, FACS, IF, IHC, IHC (fro), IHC (p), IP, WB Hôte: Lapin Polyclonal
Pubmed (340) Independent Validation (1)
N° du produit ABIN129578
$295.00
Expédition gratuite
100 μL ABIN129578
100 μL ABIN129578
local_shipping Destination: Etats-Unis
Envoi sous 1 à 2 jours ouvrables
  • Highlights
    • Frequently cited in scientific publications with 300+ citations
    • Validated inhouse for relevant applications
    • Extensive validation report for IF provided by one of your peers
    • Designed to detect RFP and its variants
    Antigène
    Reactivité
    Discosoma
    Hôte
    Lapin
    Clonalité
    Polyclonal
    Conjugué
    Cet anticorp RFP est non-conjugé
    Application
    ELISA, Flow Cytometry (FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
    Fonction
    Polyclonal RFP antibody is designed to detect RFP and its variants.
    Réactivité croisée (Details)
    Expect reactivity against RFP and its variants: mCherry, tdTomato, mBanana, mPlum, mOrange and mStrawberry. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum and purified and partially purified Red Fluorescent Protein (Discosoma).
    No reaction was observed against Human, Mouse or Rat serum proteins.
    Purification

    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.

    Preadsorption: Pre-adsorbed

    Stérilité
    Sterile filtered
    Immunogène
    The immunogen is a Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.
    Immunogen Type: Recombinant Protein
    Isotype
    IgG
  • Indications d'application
    Suggested dilutions:
    ELISA: 1:28,700 - 1:48,700
    IF: 1:200 - 1:2,000
    FACS: 1:200 - 1:2,000
    WB: 1:1,000 - 1:5,000
    IHC: 1:200 - 1:2,000
    IP: User Optimized
    Commentaires

    This antibody can be used to detect RFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated polyclonal anti-RFP used in a sandwich ELISA with unconjugated anti-RFP is well suited to titrate RFP in solution. The detection antibody conjugated to biotin is subsequently reacted with streptavidin conjugated HRP.
    Fluorochrome conjugated polyclonal anti-RFP can be used to detect RFP by immunofluorescence microscopy in cell expression systems and can detect RFP containing inserts. Significant amplification of signal is achieved using fluorochrome conjugated polyclonal anti-RFP relative to the fluorescence of RFP alone.
    For immunoblotting use either alkaline phosphatase or peroxidase conjugated polyclonal anti-RFP to detect RFP or RFP containing proteins on western blots.
    Optimal titers for applications should be determined by the researcher.

    Restrictions
    For Research Use only
  • Validation #101103 (Immunofluorescence)
    'Independent Validation' signe
    by
    Centre d'Immunologie de Marseille-Luminy
    No.
    #101103
    Date
    07.03.2017
    Antigène
    RFP
    Numéro du lot
    34945
    Application validée
    Immunofluorescence
    Contrôle positif
    CX3CL1mcherry+ murine thymus
    Contrôle négative
    CX3CL1 WT murine thymus
    Conclusion

    Passed. The RFP antibody ABIN129578 specifically labels RFP+ cells in CX3CL1mcherry+ murine thymus in immunofluorescence, consistent with the expression pattern of CX3CL1mcherry.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN1043867
    Anticorps secondaire
    donkey Fab'2 anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806)
    Full Protocol
    • Harvest thymus from 7 weeks old mice in 1X Dulbecco's phosphate buffered saline (DPBS) (Gibco Life Technologies, 14200-067).
    • Fix mouse thymus in antigen fix (Diapath, P0014) for 2h at 4?C.
    • Wash tissue in 0.1M pH7.4 phosphate buffer at for 1h at 4?C.
    • Dehydrate tissue in 30% sucrose solution at 4?C ON.
    • Snap freeze tissue in Tissue Freezing Medium (ElectroMicroscopy Science, 72592-C) at -80?C.
    • Cut blocks into 25?m sections using a cryostat (Leica, CM3050 S).
    • Transfer sections to a slide.
    • Create a hydrophobic barrier on the slide around sections with Dako pen (Dako, S2002, lot 00081640).
    • Place slide in a humidified chamber and rehydrate sections in 0.1 M TrisHCl pH7.4 for 10min at RT.
    • Gently remove buffer by tapping slide.
    • Permeabilize tissue in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, lot 015K0039) and 0.5% BSA, for 20min at RT.
    • Incubate sections with primary RFP antibody (Red Fluorescent Protein) (AA 234) (antibodies-online, ABIN129578, lot 34945) diluted 1:1000 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA for 2h at RT.
    • Incubate a no primary antibody negative control in parallel in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA.
    • Wash slides 1x 5min in 0.1M TrisHCl pH7.4.
    • Incubate sections with secondary donkey Fab'2 anti-rabbit AF647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806) diluted 1:300 in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA for 2h at RT.
    • Wash slides 1x 5min in 0.1M TrisHCl pH7.4.
    • Add approximately 10µL of Slowfade Gold antifade reagent (ThermoFisher Scientific, S36937, lot 1226836) for each section and mount cover slip.
    • Image acquisition on an LSM 880 (Zeiss), 20x magnification, 1000 resolution.
    Notes
  • Format
    Liquid
    Concentration
    1.0 mg/mL
    Buffer
    0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
    Agent conservateur
    Sodium azide
    Précaution d'utilisation
    This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Conseil sur la manipulation
    Avoid cycles of freezing and thawing.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
    Date de péremption
    12 months
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  • Antigène
    Autre désignation
    RFP
    Sujet
    Synonyms: DsRed, rDsRed, Discosoma sp. Red Fluorescent Protein, Red fluorescent protein drFP583
    Fluorescent proteins such as Discosoma Red Fluorescent Protein (DsRed) from sea anemone Discosoma sp. mushroom or green fluorescent protein (GFP) from Aequorea victoria jellyfish are widely used in research practice. Fusion RFP and GFP commonly serve as marker for gene expression and protein localization. As DsRed and GFP share only 19% identity, therefore, in general, anti-GFP antibodies do not recognize DsRed protein and vice versa. Structurally, Discosoma red fluorescent protein is similar to Aequorea green fluorescent protein in terms of its overall fold (a β-can) and chromophore-formation chemistry. However, Discosoma red fluorescent protein undergoes an additional step in the chromophore maturation and obligates tetrameric structure.
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