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 E
Western blot using Rabbit Anti-Mouse IgG Peroxidase Conjugated Antibody and Mouse Anti-GFP Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: HeLa WC Lysate+GFP protein (p/n W09-000-364 [10µg]/ p/n 000-001-215 [0.1µg]). Lane 3: HeLa WC Lysate+GFP protein [10µg/0.066µg]. Lane 4: HeLa WC Lysate+GFP protein [10µg/0.033µg]. Lane 5: HeLa Whole Cell Lysate (p/n W09-000-364) [10µg]. Primary Antibody: Anti-GFP at 1:1000 overnight at 2-8C. Secondary Antibody: Rabbit Anti-Mouse IgG HRP (p/n 610-4302) at 1:40,000 for 30mins at RT. Block: BlockOut Buffer (p/n MB-073) for 60mins at RT. Expected MW: ~27kDa.
Western Blot of Rocklands 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 B501-0500) o/n at 4 C. The membrane was then probed with a 1:2,500 dilution of HRP Rb-a-Mouse IgG [H&L] (p/n 610-4302) for 45 min at RT. FemtoMax Super Sensitive Chemiluminescent HRP Substrate (p/n FEMTOMAX-110) was added and the image was captured using a BioSpectrum Imaging System with a BioChemi 500 -28C cooled 4.0 Megapixel CCD Camera from UVP (www.uvp.com, Upland, CA). Binning: 5X5. Exposure time: 2 min. Aperture open. No emission or excitation filters used.
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 010-0102). The starting dilution of antibody was 5µg/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:185,000. 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 TMBE-1000.
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