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RFP Antibody Pre-adsorbed

Catalogue number:
600-401-379S
Size:
25 µl
Product is available in:
  • UK
  • Ireland
  • Europe
  • USA
  • Rest of World
£145.00 Estimated delivery Thu 1 Aug
Shipping is calculated in checkout
Applications:

ELISA, Flow Cytometry, Immunohistochemistry, IF Microscopy, Western Blot

Antibody Host:

Rabbit

Species Reactivity:

RFP; tdTomato

Antibody Isotype:

IgG

Antibody Type:

Polyclonal

Immunogen:

The immunogen is a Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom

Alternative Names:

rabbit anti-RFP antibody, DsRed, rDsRed, Discosoma sp. Red Fluorescent Protein, Red fluorescent protein drFP583

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Product Description:

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. Rockland offers many controls, monoclonal, and polyclonal antibodies for RFP. Polyclonal anti-RFP is designed to detect RFP and its variants. This antibody has been tested by ELISA, western blot, IF, and IHC, and is suitable for use in IP, ICC, dual RNA-FISH, iDISCO+, IEM, and FLOW. 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 (code # S000-03) . 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.

Storage Temperature:

Store vial at -20°C or below prior to opening. This vial contains a relatively low volume of reagent

Immunofluorescence Microscopy of Rabbit
Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: HopERCre/+; R26Tom/+ mice. Fixation: 0.5% PFA. Antigen retrieval: Tamoxifen. Primary antibody: RFP antibody 1:100 for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:10,000 for 45 min at RT. Staining: Hop-derived cells in the hair follicle, labeled in red.
Immunohistochemistry of Anti-RFP Antibod
Immunohistochemistry of Anti-RFP Antibody. Tissue: Mouse gut tissue in tomato transgenic mice. Fixation: formalin fixed paraffin embedded. Antigen retrieval: heat 5 min at high temp in 1X rodent decloaker. Primary antibody: RFP antibody at 1:200 for 1 h 30 min at RT. Secondary Antibody: HRP anti-rabbit (p/n 611-103-122). Image courtesy: Kwan Hyun Kim.
Immunofluorescence Microscopy of Rabbit
Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: (10X) Mouse lung tissue. Fixation: 4% PFA. Antigen retrieval: Heat. Primary antibody: Anti-RFP antibody at 1:50 for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:250 for 1 hr at RT. Staining: SPC+ cells are RFP positive in red.
Immunofluorescence Microscopy of Rabbit
Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: (10X) Mouse E14.5 embryo heart tissue. Fixation: 4% PFA. Antigen retrieval: Heat. Primary antibody: Anti-RFP antibody at 1:50 for 1 h at RT. Secondary antibody: Fluorescein rabbit secondary antibody at 1:250 for 1 hr at RT. Staining: cardiac cells are RFP positive in red in tomato transgenic mice.
Immunofluorescence Microscopy of Rabbit
Immunofluorescence Microscopy of Rabbit Anti-RFP antibody. Tissue: DsRed transgenic mouse retina. Fixation: 4% PFA. Blocking: 3% BSA, 0.3% Triton Primary antibody: RFP antibody at 1:100 for 12 h at 4°C. Secondary antibody: Alexa488 secondary antibody at 1:10,000 for 4 hours at RT. Localization: RFP is nuclear and occasionally cytoplasmic. Staining: labeled in red.
Western blot of RFP recombinant protein
Western blot of RFP recombinant protein detected with Rockland's polyclonal anti-RFP antibody. Lane 1 shows no reaction against a GFP recombinant protein present in 10 µg of HeLa cell extract. Lane 2 shows a single band detected in 10 µg of a HeLa lysate containing RFP recombinant protein as a 27 kDa band. A 4-12% Bis-Tris gradient gel (Invitrogen) was used for SDS-PAGE. The membrane was blocked and then probed with Anti-RFP diluted 1:2,500 for 1 h at RT followed by washes and reaction with a 1:5,000 dilution of IRDye™800 conjugated Goat-a-Rabbit IgG [H&L] MX (611-132-122). IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
Western Blot of RFP Antibody Pre-Absorbe
Western Blot of RFP Antibody Pre-Absorbed. Lane 1: RFP. Lane 2: Human IgG. Lane 3: Goat IgG. Lane 4: Mouse IgG. Load: 50ng per lane. Primary antibody: RFP Antibody Pre-Absorbed at 1:1,000 overnight at 4°C. Secondary antibody: Peroxidase conjugated rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: MB-070 Blocking Buffer for 30 min at RT. Predicted/Observed size: 27 kDa, 30 kDa.
Western Blot of Rabbit anti-RFP Antibody
Western Blot of Rabbit anti-RFP Antibody. Lane 1: Opal Prestained Marker (p/n MB-210-0500). Lane 2: 50ng of RFP. Primary Antibody: Anti-RFP at 1µg/mL overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit HRP (p/n 611-103-122) at 1:70,000 for 30mins at RT. Block: BlockOut Universal blocking buffer (p/n MB-073). Expect ~27kDa.
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