Mouse IgG (H&L) Secondary Antibody Peroxidase Conjugated

• Catalog Number: orb347544
• Category: Antibodies
• Description: Mouse IgG (H&L) antibody (Peroxidase)
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Mouse
• Isotype: IgG
• Immunogen: Mouse IgG whole molecule
• Concentration: 2.0 mg/mL
• Dilution range: ELISA: 1:50,000 - 1:300,000, IHC: 1:500 - 1:2,500, WB: 1:10,000 - 1:30,000
• Form/Appearance: Lyophilized
• Purity: HRP Secondary Antibody Conjugate was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Rabbit Serum, Mouse IgG and Mouse Serum.
• Conjugation: HRP
• Storage: Store mouse secondary antibody conjugate at 4° C prior to restoration. For extended storage aliquot antibody 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-Mouse IgG Secondary Antibody Peroxidas
• Note: For research use only
• Application notes: Anti-Mouse secondary antibody conjugated to horseradish peroxidase (HRP) generated in rabbit detects specifically Mouse IgG whole molecule. Anti-Mouse IgG peroxidase antibody has been tested by ELISA and western blot and is suitable for ELISA, Sandwich ELISA, titration assays, western-blot, immunoprecipitation, Immunohistochemistry as well as other HRP antibody based assays. Specific conditions for reactivity and signal detection should be optimized by the end user.
• Expiration Date: 12 months from date of receipt.

ELISA Results of Rabbit Anti-Mouse IgG Peroxidase Antibody tested against purified Mouse IgG protein. Each well was coated in duplicate with 10 µg of Mouse IgG (p/n orb2652749). The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:185000. 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 Biorbyt's Anti-Mouse IgG Antibody Peroxidase Conjugated detecting mouse IgG. Mouse IgG (1.0 mg) was separated on a 4-20% gradient gel and then transferred to Whatman Protran BA 85 nitrocellulose (0.45 µm pore size), and blocked with 5% BLOTTO (p/n orb348624) o/n at 4°C. The membrane was then probed with a 1:2500 dilution of HRP Rb-a-Mouse IgG [H&L] (p/n orb347544) for 45 min at RT.

Western blot using Rabbit Anti-Mouse IgG Peroxidase Conjugated Antibody and Mouse Anti-GFP Antibody. Lane 1: Opal Prestained Molecular Weight Marker. Lane 2: HeLa WC Lysate + GFP protein (p/n orb348668 [10 µg]/ p/n orb345957 [0.1 µg]). Lane 3: HeLa WC Lysate + GFP protein [10 µg/0.066 µg]. Lane 4: HeLa WC Lysate + GFP protein [10 µg/0.033 µg]. Lane 5: HeLa Whole Cell Lysate (p/n orb348668) [10 µg]. Primary Antibody: Anti-GFP at 1:1000 overnight at 2-8°C. Secondary Antibody: Rabbit Anti-Mouse IgG HRP (p/n orb347544) at 1:40000 for 30 mins at RT. Block: BlockOut Buffer (p/n orb348644) for 60 mins at RT. Expected MW: ~27kDa.