RAT IgG (H&L) antibody (Peroxidase)

• Catalog Number: orb347858
• Category: Antibodies
• Description: RAT IgG (H&L) antibody (Peroxidase)
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Rat
• Isotype: IgG
• Immunogen: Rat IgG whole molecule
• Concentration: 2.0 mg/mL
• Dilution range: ELISA: 1:170,000, IHC: 1:500 - 1:2,500, WB: 1:1,000 - 1:5,000
• Form/Appearance: Lyophilized
• Purity: Secondary Antibody Conjugated was prepared from monospecific antiserum by immunoaffinity chromatography using Rat 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-Peroxidase, anti-Rabbit Serum, Rat IgG and Rat Serum.
• Conjugation: HRP
• Storage: Store secondary antibody 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.
• Buffer/Preservatives: 0.01% (w/v) Gentamicin Sulfate. Do NOT add Sodium Azide!
• Alternative names: Rabbit anti-Rat IgG peroxidase Conjugated Antibody
• Note: For research use only
• Application notes: Anti-Rat IgG Peroxidase conjugated antibody has been tested by ELISA and is suitable for western blot, ELISA, immunoperoxidase electron microscopy and immunohistochemistry as well as other peroxidase-antibody based enzymatic assays requiring lot-to-lot consistency.
• Expiration Date: 12 months from date of receipt.

ELISA results of Rabbit Anti-Rat IgG Antibody Peroxidase Conjugated tested against purified Rat IgG protein. Each well was coated in duplicate with 1.0 µg of Rat IgG [Green Line]. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:41800. 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.

Western Blot of Anti-Rat IgG (H&L) (RABBIT) Antibody (p/n orb347848). Lane M: 3 μl Molecular Ladder. Lane 1: Rat IgG whole molecule. Lane 2: Rat IgG F(c) Fragment (p/n orb346338). Lane 3: Rat IgG Fab Fragment (p/n orb346340). Lane 4: Rat IgM Whole Molecule (p/n orb346341). All samples were reduced. Load: 50 ng per lane. Block: orb348637 for 30 min at RT. Primary Antibody: Anti-Rat IgG (H&L) (RABBIT) Antibody (p/n orb347848) 1:1000 for 60 min at RT. Secondary Antibody: Anti-Rabbit IgG (GOAT) Peroxidase Conjugated Antibody (p/n orb347654) 1:40000 in orb348637 for 30 min at RT. Predicted/Observed Size: 25 and 55 kDa for Rat IgG, 25 kDa for F(c) and Fab, 78 and 25 kDa for IgM. Rat F(c) migrates slightly higher.