Rabbit IgG (H&L) Antibody Alkaline Phosphatase Conjugated

• Catalog Number: orb347681
• Category: Antibodies
• Description: RABBIT IgG (H&L) antibody (Alkaline Phosphatase)
• Species/Host: Goat
• Clonality: Polyclonal
• Tested applications: DOT, ELISA, IHC, WB
• Reactivity: Rabbit
• Isotype: IgG
• Immunogen: Anti-Rabbit IgG (H&L) was produced by repeated immunization with rabbit whole IgG molecule in goat.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:3,000 - 1:25,000, IHC: 1:200 - 1:1,000, WB: 1:1,000 - 1:4,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-Rabbit IgG (H&L) Antibody Alkaline Phosphatase conjugated was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Alkaline Phosphatase (calf intestine), anti-Goat Serum, Rabbit IgG and Rabbit Serum.
• Conjugation: AP
• Storage: Store vial at 4° C before opening. DO NOT FREEZE. This product is stable at 4° C as an undiluted liquid. Dilute only prior to immediate use. Freezing alkaline phosphatase conjugates will result in a substantial loss of enzymatic activity.
• Buffer/Preservatives: 0.1% (w/v) Sodium Azide
• Alternative names: goat anti-Rabbit IgG Antibody Alkaline Phosphatase
• Note: For research use only
• Application notes: Anti-Rabbit IgG (H&L) alkaline phosphatase conjugated antibody has been tested by ELISA, dot blot, and western blot and is suitable for immunoblotting (western or dot blot), ELISA, immunohistochemistry as well as other alkaline phosphatase-antibody based enzymatic assays requiring lot-to-lot consistency.
• Expiration Date: 12 months from date of receipt.

Dot Blot of Rabbit IgG Antibody Alkaline Phosphatase Conjugated. Antigen: Rabbit IgG. Load: Lane 1 - 200 ng, Lane 2 - 66.7 ng, Lane 3 - 22.2 ng, Lane 4 - 7.4 ng, Lane 5 - 2.5 ng. Primary antibody: none. Secondary antibody: Rabbit IgG Antibody Alkaline Phosphatase Conjugated at 1:1000 for 60 min at RT. Block: orb348637 for 60 min at RT. Reaction visualized using alkaline phosphatase substrate for 30 seconds at RT.

ELISA results of Goat Anti-Rabbit IgG Antibody Alkaline Phosphatase Conjugated tested against purified Rabbit IgG protein. Each well was coated in duplicate with 10 µg of Rabbit IgG (p/n orb2652747). The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:45700. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gelatin as blocking buffer and TMB substrate p/n orb348651.

Typical experiment image where the complexes Ag–AbI–AbII are attached to the nanoparticles trapped in the magnetic traps and the fluorogenic substrate is flowed at different rates, where the measurement areas selected for the fluorescence analysis are shown. Measurement with a flow rate of (a) 1 µl/h, (b) 2 µl/h, (c) 5 µl/h, and (d) 10 µl/h. Experiment with anti-rabbit IgG labeled alkaline phosphatase was used (p/n orb347681) and an anti-biotin rabbit IgG (p/n orb750609) [AbI] concentration of 100 pg/ml.

Typical experiment image where the complexes Ag–AbI–AbII are attached to the nanoparticles trapped in the magnetic traps and the fluorogenic substrate is flowed at different rates, where the measurement areas selected for the fluorescence analysis are shown. Measurement with a flow rate of (a) 1 µl/h, (b) 2 µl/h, (c) 5 µl/h, and (d) 10 µl/h. Experiment with anti-rabbit IgG labeled alkaline phosphatase was used (p/n orb347681) and an anti-biotin rabbit IgG (p/n orb750609) [AbI] concentration of 100 pg/ml.

Western Blot of Goat anti-Rabbit IgG Antibody Alkaline Phosphatase Conjugated. Lane 1: Rabbit IgG. Lane 2: None. Load: 100 ng per lane. Primary Antibody: None. Secondary antibody: Alkaline Phosphatase goat secondary antibody at 1:1000 for 60 min at RT. Block: orb348637 for 30 min at RT. Predicted/Observed size: 55 and 28 kDa, 55 kDa for Rabbit IgG. Other Band(s): None.