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RFP antibody (Red Fluorescent Protein) (Atto 565) Primary Antibody

RFP Reactivity: Discosoma FACS, ICC, IF Host: Camelid (Camelidae) Monoclonal 2B12
Independent Validation (1)
Catalog No. ABIN4368323
$528.00
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200 μL
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  • Target
    Antibody Type
    Recombinant Antibody
    Reactivity
    Discosoma
    Host
    Camelid (Camelidae)
    Clonality
    Monoclonal
    Conjugate
    This RFP antibody is conjugated to Atto 565
    Application
    Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF)
    Purpose
    Camelid sdAb anti-RFP conjugated with Atto565, Clone 2B12
    Specificity
    Recognizes most common red fluorescent proteins like mRFP and derivatives like mCherry, mScarlet-i, tdTomato, dsRed and mOrange.
    Cross-Reactivity (Details)
    Does not cross-react with GFP or mTagBFP derivatives.
    Characteristics
    A dye conjugated alpaca single-domain antibody (sdAb), also referred to as VHH or nanobody. It is in the range of 15 kDa and 3 nm in molecular weight and size respectively. This means that sdAbs are ten times lighter and up to 5x smaller than a conventional IgG molecule. They can position a fluorophore up to 20 nm closer to the intended target than using conventional primary-secondary antibody complex detection.
    Immunogen
    RFP
    Clone
    2B12
    Fragment
    single-domain Antibody (sdAb)
    Labeling Ratio
    a single fluorophore is coupled to exactly one sdAb
    Featured
    Discover our best selling RFP Primary Antibody
  • Application Notes
    Recommended dilution 1:500
    Comment

    Each fluorophore is coupled to exactly one sdAb, which in turn binds to its target molecule in a monovalent fashion. The high binding affinity and a high coupling efficiency of > 95% guarantees a highly linear relation between target molecule number and fluorescent intensity. This enables you to directly count your target molecule of interest. The fluorophore is located exceptionally close to the recognized epitope (< 1.5 nm), which is ideal for all microscopy techniques.

    Restrictions
    For Research Use only
  • Validation #101078 (Immunofluorescence)
    'Independent Validation' Badge
    by
    Centre d'Immunologie de Marseille-Luminy
    No.
    #101078
    Date
    03/07/2017
    Antigen
    RFP
    Lot Number
    1602
    Method validated
    Immunofluorescence
    Positive Control
    CX3CL1mcherry+ murine thymus
    Negative Control
    CX3CL1 WT murine thymus
    Notes

    Passed. The RFP (Atto 565) antibody ABIN4368323 specifically labels RFP+ cells in CX3CL1mcherry+ murine thymus in immunofluorescence, consistent with the expression pattern of CX3CL1mcherry.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN4368323
    Secondary Antibody
    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 0.1M TrisHCl pH7.4 for 10min at RT.
    • Gently remove buffer by tapping the 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 camelid anti-RFP antibody (Atto 565) (antibodies-online, ABIN4368323, lot 1602) diluted 1:1000 0.1M TrisHCl pH7.4 containing 2% Triton X-100 and 0.5% BSA for 2h at RT.
    • Wash slides 1x 5min in 0.1M TrisHCl pH7.4.
    • Add approximately 10?L 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.
    Experimental Notes
  • Format
    Liquid
    Concentration
    5 μM
    Buffer
    5 μM fluorescently labeled sdAb in buffered saline, 50 % glycerol, 0.09 % sodium azide.
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handling Advice
    Protect from light!
    Storage
    -20 °C,-80 °C
    Storage Comment
    Up to 3 months store at -20 °C. Up to 12 months store at -80 °C or below
    Expiry Date
    12 months
  • Target
    Alternative Name
    RFP
    Molecular Weight
    26 kDa
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