Anti IgG (H&L)

Rockland secondary antibodies detect rabbit primary antibodies in a variety of platforms. Shown here is a serial 1:1 dilution of control rabbit IgG protein (011 0102, 250ng starting total load) co incubated with Rockland HRP conjugated Goat anti Rabbit IgG (611 103 122) and Dylight 649 conjugated goat anti Rabbit (611 143 122) 1:20K in MB 070. Blot was dried and imaged (A) on Biorad Versa Doc (30 sec, DyLight649), (B) LiCor Odyssey Reader (700 nm), (C ) Rewetted incubated with Femtomax 110 reimaged using BioVersaDoc (for 60 sec), (D) Incubated with TMB substrate TMBM for 5 minutes and scanned, and (E) Rewetted for Chemiluminescence and imaged for 90 sec on the BioRad VersaDoc Imager,

HRP-conjugated Goat-Anti-Rabbit (p/n 611-103-122) secondary antibody was used at 1:40,000 in MB-070 blocking buffer to detect a rabbit primary antibody by Western Blot. Anti-p27 antibody (200-401-C30, 1:1000 RT 30 minutes) showed detection of 0.1 ug of recombinant p27 protein. Lane 1: Molecular weight markers. Lane 2: MBP-p27 fusion protein (arrow; expected MW: 73.3 kDa). Lane 3: MBP alone. Protein was run on a 4-20% gel, then transferred to 0.45 um nitrocellulose and blocked with 1% BSA-TTBS (p/n MB-013, diluted to 1X) overnight at 4C. Blot was imaged on the VersaDoc MP 4000 imaging system (Bio-Rad).

ELISA Results of Goat Anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt, Sh Serum Proteins) tested against purified Goat Anti-Rabbit IgG Antibody HRP Conjugated MX10. Each well was coated in duplicate with 10 ug of Rabbit IgG (p/n 011-0102) [Red Line], Bovine IgG (p/n 001-0102) [Green Line], Chicken IgG (p/n 003-0102) [Blue Line], Goat IgG (p/n 005-0102) [Purple Line], Guinea Pig, Hamster, Horse, Human, Mouse, Rat, and Sheep tested with similar results. The working dilution for Rabbit IgG is 1:109,000. The starting dilution of antibody was 5μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% Fish Gel/PBS Blocking buffer (p/n MB-066), and TMB substrate (p/n TMBE-1000).

Western Blot of Unconjugated Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n 611-101-122). 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 per lane. Block: MB-070 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n 611-101-122) 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody (p/n CUST10) 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.