Rabbit IgG (H&L) Secondary Antibody Fluorescein Conjugated

• Catalog Number: orb347667
• Category: Antibodies
• Description: Rabbit IgG (H&L) antibody (FITC)
• Species/Host: Goat
• Clonality: Polyclonal
• Tested applications: DOT, FC, FLISA, IF, WB
• Reactivity: Rabbit
• Isotype: IgG
• Immunogen: Rabbit IgG whole molecule
• Antibody Type: Secondary Antibody
• Concentration: 1.0 mg/mL
• Dilution range: FLISA: 1:10,000 - 1:50,000, FC: 1:500 - 1:2,500, IF: 1:1,000 - 1:5,000
• Form/Appearance: Lyophilized
• Purity: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit 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-Fluorescein, anti-Goat Serum, Rabbit IgG and Rabbit Serum.
• Conjugation: FITC
• Storage: Store vial 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) Sodium Azide
• Alternative names: Goat anti-Rabbit IgG Antibody fluorescein Conjugat
• Note: For research use only
• Application notes: Anti-Rabbit IgG Antibody Fluorescein has been tested by dot blot and western blot and is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.
• Expiration Date: 12 months from date of receipt.

Biorbyt FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody was used to detect nanogram – picogram levels of rabbit IgG by dot blot on nitrocellulose membrane. 4 ul each of serial 1 in 4 dilutions of rabbit IgG were dotted on nitrocellulose and allowed to dry. Membrane was blocked in 3% BSA for 10 minutes dried for later use and rewetted with orb348637. Blot was incubated in Biorbyt fluorescein conjugated goat anti rabbit orb347666 1:10000 and Biorbyt HRP conjugated goat anti Rabbit orb347673 1:10000, dried and: A. Blot was imaged on the BioRad VersaDoc with filter settings appropriate for Fluorescein/DyLight 488. B. Blot was rewetted with TBS, incubated with FEMTOMAX chemiluminescent substrate for 1-3 minutes and imaged for 60sec on the BioRad VersaDoc Imaging System. C. Blot was rinsed with TBS and DIH2O, incubated for 5 minutes with Biorbyt TMB Substrate for Western Blot MaxTag (1 ml of TMBM-102 + ~9 ml of orb342326), dried overnight.

Western Blot of Anti-Rabbit IgG (H&L) (GOAT) Antibody (p/n orb347660). Lane M: 3 µl Molecular Ladder. Lane 1: Rabbit IgG whole molecule (p/n orb2652747). Lane 2: Rabbit IgG F(ab) Fragment (p/n orb2652745). Lane 3: Rabbit IgG F(c) Fragment (p/n orb346280). Lane 4: Rabbit IgM Whole Molecule (p/n orb346313). Lane 5: Normal Rabbit Serum. All samples were reduced. Load: 50 ng per lane. Block: orb348637 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody (p/n orb347660) 1:1000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40000 in orb348637 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.

Western blot of Fluorescein conjugated Goat Anti-Rabbit IgG secondary antibody. Lane 1: Rabbit IgG. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Fluorescein goat secondary antibody at 1:1000 for 60 min at RT. Blocking: orb348637 for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Rabbit IgG. Other band(s): None.