Recombinant RFP antibody

• Catalog Number: orb1463559
• Category: Antibodies
• Description: Recombinant RFP antibody
• Clonality: Recombinant
• Clone Number: RFP40
• Tested applications: DOT, ELISA, WB
• Reactivity: Other
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:31,000, WB: 1:1000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-RFP (VHH) Antibody is a recombinant antibody. The clone was isolated from a library prepared from a hyper-immunized llama host and purified by affinity chromatography from bacterial culture. Expect reactivity against RFP-labeled probes and proteins.
• Conjugation: Unconjugated
• UniProt ID: Q9U6Y8
• Source: E. Coli
• Biological Origin: E. Coli
• Storage: Store at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: recombinant anti-RFP antibody, Red Fluorescent Pro
• Note: For research use only
• Application notes: Anti-RFP is a his-tagged monoclonal recombinant antibody designed to detect Red Fluorescent Protein and its conjugates. This antibody has been tested by dot blot and ELISA and is intended for use in immunological assays including ELISA, immunofluorescence and fluorescence activated cell sorting (FACS). The antibody can be labeled with dyes, enzymes or fluorescence, directly or secondarily, for visualization and detection of RFP-conjugated molecules by immunofluorescence. Secondary detection can be achieved using conjugated anti-His tag or anti-VHH antibodies. Optimal titers for applications should be confirmed by the end user. This antibody is not suitable for Western blot detection of denatured RFP.
• Expiration Date: 12 months from date of receipt.

Dot Blot of Anti-RFP Single Domain Antibody. Row A: Recombinant RFP protein (p/n orb345960). Amount Dotted: (1) 200, (2) 100, (3) 50, (4) 25, (5) 12.5 ng. Primary Antibody: Anti-RFP (VhH) 1.0 µg/mL for 1.0 hr at RT. Secondary Antibody: Mouse Anti-Flag Peroxidase (p/n orb344540) at 1:5000 for 30 mins at RT. Block: BlockOut Buffer (p/n orb348644). Exposure: 10 seconds.

ELISA Results of purified recombinant Anti-RFP Single Domain Antibody reactive against RFP. The working dilution for clone RFP40 is 1:31000. Each well was coated in duplicate with 1.0 µg of RFP (p/n orb345960). The starting dilution of antibody was 45 µg/mL and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% Fish Gel/PBS blocking buffer, Mouse Anti-FLAG Tag HRP conjugated Antibody (p/n orb344540) and TMB substrate (p/n orb348651).

Western Blot Results of Anti-RFP (VHH) Single Domain Antibody. Lane 1: Opal Prestained Molecular Weight. Lane 2: HEK293 Whole Cell Lysate (p/n orb348669) [10 ug] (-). Lane 3: HEK293 Whole Cell Lysate (p/n orb348669) [10 ug] + GFP rec. protein (p/n orb345957) [1 ug] (-). Lane 4: HEK293 Whole Cell Lysate (p/n orb348669) [10 ug] + RFP rec. protein (p/n orb345960) [1 ug] (+). Lane 5: RFP rec. protein (p/n orb345960) [1 ug] (+). Primary Antibody: Anti-RFP (VHH) Single Domain Antibody (p/n orb1463559) at 1:1000 for overnight at 4°C. Secondary Antibody: Mouse Anti-6x His DL649 conjugated at 1:10000 for 1 hr at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n orb348637) overnight at 4°C. Expected MW: ~27 kDa for RFP protein.