Luciferase antibody

• Catalog Number: orb344220
• Category: Antibodies
• Description: Luciferase antibody
• Species/Host: Goat
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC, WB
• Isotype: IgG
• Immunogen: Luciferase [Photinus pyralis (Firefly)]
• Concentration: 1.0 mg/ml
• Dilution range: ELISA: 1:200 - 1:1000, IHC: 1:1500, IF: User Optimized, WB: 1:1000 - 1:5000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum as well as purified and partially purified Luciferase [Photinus pyralis (Firefly)]. No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.
• Conjugation: Unconjugated
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: goat anti-Luciferase Antibody, Luciferin 4-monooxy
• Note: For research use only
• Application notes: Anti-Luciferase Antibody has been tested in Western Blot, IHC, IF, and ELISA. Expect a band ~60kDa in appropriate cell lysates. Although not tested, this antibody would be useful in immunoprecipitation, immunocytochemistry, and most immunological methods requiring high titers and specificity.
• Expiration Date: 12 months from date of receipt.

ELISA Results of Goat Anti-Luciferase Antibody. Each well was coated in duplicate with 1.0 µg of Luciferase [Firefly]. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:6050. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using TMB substrate p/n orb348651.

Western Blot of Goat anti-Luciferase antibody. Lane 1 Marker: Opal Pre-stained ladder. Lane 2: HEK293 lysate (p/n orb348669). Lane 3: HeLa Lysate (p/n orb348668). Lane 4: CHO/K1 Lysate. Lane 5: E-coli HCP Control (p/n orb342262). Lane 6: J774A.1 Lysate. Lane 7: C2C12 Lysate (p/n orb348725). Lane 8: Mouse Embryonic Fibroblast Lysate (p/n orb348722). Lane 9: NIH/3T3 Lysate (p/n orb348714). Lane 10: Mouse Liver Lysate (p/n orb348718). Lane 11: PC-12 Lysate (p/n orb348733). Lane 12: Rat Brain Lysate (p/n orb348727). Lane 13: Rat Testis Lysate. Lane 14: Luciferase [Photinus pyralis (Firefly)]. Lane 15 Marker: Opal Pre-stained ladder. Load: 10 µg of lysate or 50 ng of purified protein per lane. Primary antibody: Luciferase antibody at 1:1000 overnight at 4C. Secondary antibody: Peroxidase Goat secondary antibody (p/n orb347032) at 1:40000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted/Observed size: ~60 kDa for Luciferase.

Western Blot of Goat anti-Luciferase Antibody. Lane 1: Luciferase. Load: 50 ng per lane. Primary antibody: Luciferase antibody at 1:1000 overnight at 4°C. Secondary antibody: Peroxidase goat secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: (p/n orb348637) for 30 min at RT. Predicted/Observed size: ~60 kDa for Luciferase. Other band(s): None.