0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Form:
Lyophilized
Target:
Rabbit
Antibody Type:
Secondary Antibody
Application Dilute:
FLISA: >1:20,000, IF Microscopy: >1:5,000, WB: >1:10,000
Application Notes:
Anti-Rabbit IgG (H&L) DyLight 680 Antibody has been tested by dot blot and is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, incl
DyLight(TM) dyes can be used for two-color Western Blot detection with low background and high signal. Anti-tubulin was detected using a DyLight(TM) 680 conjugate. Anti-TNFa was detected using a DyLight(TM) 800 conjugate. The image was captured using the Odyssey Infrared Imaging System developed by LI-COR.
Western Blot of Anti-Rabbit IgG (H&L) (DONKEY) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n 611-701-127). Lane M: 3 µl Molecular Ladder. Lane 1: Rabbit IgG whole molecule (p/n 011-0102). Lane 2: Rabbit IgG F(ab) Fragment (p/n 011-0105). Lane 3: Rabbit IgG F(c) Fragment (p/n 010-0103). Lane 4: Rabbit IgM Whole Molecule (p/n 011-0107). Lane 5: Normal Rabbit Serum (p/n B309). All samples were reduced. Load: 50 ng of IgG, F(ab), F(c) and Serum, 25 ng of IgM. Block: MB-070 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (DONKEY) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n 611-701-127) 1:7,500 for 60 min at RT. Secondary antibody: Anti-Donkey IgG (GOAT) Peroxidase Conjugated Antibody (p/n 616-1302) 1:40,000 in MB-070 for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM. Rabbit F(c) migrates slightly higher.
Properties of DyLight(TM) Conjugates.
DyLight(TM) 680 Fluorescence Spectra.
Dot Blot results of Donkey Anti-Rabbit IgG Antibody DyLight(TM)680 Conjugate. Dots are Rabbit IgG: (1) 100ng, (2) 33.3ng, (3) 11.1ng, (4) 3.70ng, (5) 1.23ng. Primary Antibody: none. Secondary Antibody: Donkey Anti-Rabbit IgG Antibody DyLight(TM)680 at 1ug/mL in MB-070 1hr RT. Imaged with BioRad ChemiDoc, DyLight(TM)680 filter.
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