RFP anticorps (Red Fluorescent Protein) (AA 234)

Details for Product anti-RFP Antibody No. ABIN1043867, Fournisseur: Connectez-vous pour afficher
Antigène
Épitope
AA 234
30
6
3
2
1
1
Reactivité
Discosoma
81
50
36
10
7
2
1
1
Hôte
Lapin
96
37
14
9
1
Clonalité
Polyclonal
Conjugué
Cet anticorp RFP est non-conjugé
9
7
6
3
3
3
3
3
3
3
3
3
3
3
2
2
2
2
2
2
2
1
1
1
1
1
1
Application
Fluorescence Microscopy (FM), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
133
70
61
53
32
29
7
4
4
3
2
1
1
1
1
1
1
Options
Fournisseur
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Supplier Product No.
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'Independent Validation' signe
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 ABIN1043867 specifically labels RFP+ cells in CX3CL1mcherry+ murine thymus in immunofluorescence, consistent with the expression pattern of CX3CL1mcherry.

Anticorps primaire ABIN1043867
Anticorps secondaire donkey Fab'2 anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806)
Protocole
  • 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, ABIN4368323, lot 1602) 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 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) (AA 234) antibody (ABIN1043867)

CX3CL1 positive cells in CX3CL1mcherry murine thymus (A, mCherry) were stained wit...

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
Specificité This product was prepared from monospecific antiserum by imunoaffinity chromatography using Red Fluorescent Protein (Discosoma) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Expect reactivity against RFP and its variants: mCherry, tdTomato, mBanana, mOrange, 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. ELISA was used to confirm specificity at less than 0.1% of target signal.
Attributs du produit Antibodies to RFP (Discosoma spp.) are intended for use in immunological assays including ELISA, western blotting, fluorometry and fluorescence activated cell sorting (FACS).
Stérilité Sterile filtered
Autre désignation RFP (RFP Antibody Extrait)
Sujet Antibodies to RFP (Discosoma spp.) are intended for use in immunological assays including ELISA, western blotting, fluorometry and fluorescence activated cell sorting (FACS).
Synonyms: DsRed, rDsRed, Discosoma sp. Red Fluorescent Protein, Red fluorescent protein drFP583
Domaine de recherche Tags/Labels
Indications d'application Suggested dilutions:
ELISA : 1:90000
IF Microscopy : 1:200-1:2000
Western Blot : 1:2000 - 1:10000
Immunochemistry: 1:200-1:2000
Other Dilution: User Optimized
Commentaires

Gene Name: DsRed

Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
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.
Stock 4 °C/-20 °C
Stockage commentaire Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. Avoid cycles of freezing and thawing. 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. Expiration date is one (1) year from date of opening.
Date de péremption 12 months
Supplier Images
Western Blotting (WB) image for anti-Red Fluorescent Protein (RFP) (AA 234) antibody (ABIN1043867) Western blot of RFP recombinant protein detected with polyclonal anti-RFP antibody. L...
Produit citée dans: Viganò, Möbius, Götz, Dimou: "Transplantation reveals regional differences in oligodendrocyte differentiation in the adult brain." dans: Nature neuroscience, Vol. 16, Issue 10, pp. 1370-2, 2013 (PubMed). (1:500).

Gross, Baird, Hoffman, Baldridge, Tsien: "The structure of the chromophore within DsRed, a red fluorescent protein from coral." dans: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, Issue 22, pp. 11990-5, 2000 (PubMed).

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