RFP antibody (Red Fluorescent Protein) (AA 234)

Details for Product anti-RFP Antibody No. ABIN1043867, Supplier: Log in to see
Antigen
Epitope
AA 234
30
6
3
2
2
1
Reactivity
Discosoma
82
50
36
10
7
2
1
1
Host
Rabbit
96
38
14
9
1
Clonality
Polyclonal
Conjugate
This RFP antibody is un-conjugated
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
71
61
53
32
29
7
4
4
3
2
1
1
1
1
1
1
Options
Supplier
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'Independent Validation' Badge
Antigen RFP
Lot Number 34945
Method validated Immunofluorescence
Positive Control CX3CL1mcherry+ murine thymus
Negative Control CX3CL1 WT murine thymus
Notes

Passed. The RFP antibody ABIN1043867 specifically labels RFP+ cells in CX3CL1mcherry+ murine thymus in immunofluorescence, consistent with the expression pattern of CX3CL1mcherry.

Primary Antibody ABIN1043867
Secondary Antibody donkey Fab'2 anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806)
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, 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...

Immunogen 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
Specificity 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.
Characteristics Antibodies to RFP (Discosoma spp.) are intended for use in immunological assays including ELISA, western blotting, fluorometry and fluorescence activated cell sorting (FACS).
Sterility Sterile filtered
Alternative Name RFP (RFP Antibody Abstract)
Background 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
Research Area Tags/Labels
Application Notes 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
Comment

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
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C/-20 °C
Storage Comment 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.
Expiry Date 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...
Product cited in: Viganò, Möbius, Götz, Dimou: "Transplantation reveals regional differences in oligodendrocyte differentiation in the adult brain." in: 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." in: 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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