RFP antibody (Red Fluorescent Protein) (Atto 565)

Details for Product anti-RFP Antibody No. ABIN4368323, Supplier: Log in to see
Antigen
Reactivity
Discosoma
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50
36
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2
1
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Host
Camelid (Camelidae)
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39
13
9
1
Clonality (Clone)
Monoclonal ()
Conjugate
This RFP antibody is conjugated to Atto 565
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3
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3
3
3
2
2
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2
2
2
1
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1
Application
Immunocytochemistry (ICC), Immunofluorescence (IF)
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7
5
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Options
Supplier
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Supplier Product No.
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'Independent Validation' Badge
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.

Primary Antibody ABIN4368323
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.
Validation Images
Immunofluorescence validation image for anti-Red Fluorescent Protein (RFP) antibody (Atto 565) (ABIN4368323)

A. Unstained CX3CL1 positive cells in CX3CL1mcherry+ murine thymus. B. ...

Purpose FluoTag-Q series - high affinity monovalent binders designed for quantitative imaging
Brand FluoTag-Q
Immunogen RFP
Clone 2B12
Fragment VHH fragment
Specificity Most common red fluorescent proteins like mRFP, mCherry, dsRed, tdTomato
Cross-Reactivity (Details) Does not cross-react with GFP, mTagBFP or mTagRFP or their most common derivatives
Characteristics FluoTag-Q anti-RFP is derived from an in-house developed single-domain antibody (sdAb) recognizing the most common red fluorescent proteins like mRFP, mCherry, DsRed and other DsRed derivatives with high affinity and specificity. In our FluoTag-Q series, each fluorophore is coupled to exactly one FluoTag, 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.
Alternative Name RFP (RFP Antibody Abstract)
Background Red fluorescent protein RFP and its derivates have become very popular and universal tools in cell biology. It is a monomeric protein and an excellent fusion partner for chimeric proteins. The possibility to fuse RFP to the N- or the C-terminus of the protein of interest enables a wide range of application. However for some microscopy technics (like STROM or STED) more photostable or blinking fluorophores are needed, but also in conventional immnuofluorescence studies an amplification of the signal or the possible to image at another wavelength might be preferable. With our FluoTags the potential of RFP fusion proteins can be exceeded by far.
Molecular Weight 26 kDa
Research Area Tags/Labels
Application Notes 1:1000 (IF, ICC)
Restrictions For Research Use only
Format Liquid
Concentration 5 μM
Buffer PBS, 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
Storage Comment Up to 3 months: -20 °C Up to 12 months: -80 °C or below
Supplier Images
Immunofluorescence (IF) image for anti-Red Fluorescent Protein (RFP) antibody (Atto 565) (ABIN4368323) 3T3 cells transfected with mCherry-tubulin and stained with FluoTag-Q anti-RFP Atto 5...
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