Rabbit IgG Fc Antibody Pre-Adsorbed

• Catalog Number: orb347664
• Category: Antibodies
• Description: Rabbit IgG F(c) antibody
• Species/Host: Goat
• Clonality: Polyclonal
• Tested applications: DOT, ELISA, IHC, WB
• Reactivity: Rabbit
• Isotype: IgG
• Immunogen: Anti Rabbit IgG F(c) was produced by repeated immunization with rabbit IgG F(c) fragment in goat.
• Concentration: 2mg/mL
• Dilution range: ELISA: 1:20,000 - 1:100,000, IHC: 1:1,000 - 1:5,000, WB: 1:2,000 - 1:10,000
• Form/Appearance: Liquid (sterile filtered)
• 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-Goat Serum, Rabbit IgG, Rabbit IgG F(c) and Rabbit Serum. No reaction was observed against Rabbit IgG F(ab) or Human Serum Proteins.
• Conjugation: Unconjugated
• Storage: Store vial at 4° C prior to opening. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: goat anti-Rabbit F(c) Antibody, Gt-a-Rabbit IgG F(
• Note: For research use only
• Application notes: Anti-Rabbit IgG F(c) is tested by immunoelectrophoresis, ELISA, dot blot, and western blot. This product is suitable for use in western-blot, competitive western-blot, and competitive ELISA assays. Specific conditions for reactivity and signal detection should be optimized by the end user. Expect ~25kDa for Rabbit IgG F(c) Fragment.
• Expiration Date: 12 months from date of receipt.

Dot Blot of Goat Anti-RABBIT IgG F(c) Antibody (Min X Human Serum Proteins). Row A: Rabbit IgG whole Molecule (p/n orb2652747). Row B: Rabbit F(c) (p/n orb346310). Row C; Rabbit F(ab). Samples Loaded: 1- 100 ng, 2- 33.33 ng, 3- 11.11 ng, 4- 3.70 ng, 5- 1.23 ng. Primary Antibody: Goat Anti-RABBIT IgG F(c) Mx Hu at 1.0 µg/ml for 60 mins at RT. Secondary Antibody: Custom HRP at 1:40000 for 30 mins at RT. Blocking (p/n orb348637) for 60 mins at RT. Exposure: 5 sec.

Dot Blot Results of Goat Anti-Rabbit F(c) mx Human IgG. Row A: Rabbit F(c) protein (p/n orb346310). Row B: Rabbit F(ab) protein (p/n orb2652745). Row C: Human IgG protein (orb346219). Amount Dotted: (1) 10 ng, (2) 3.33 ng, (3) 1.11 ng, (4) 0.37 ng, (5) 0.12 ng. Primary Antibody: Anti-Rabbit F(c) at 1.0 µl/mL for 1hr at RT. Secondary Antibody: Donkey Anti-Goat IgG Peroxidase Conjugate at 1:40000 for 30 mins at RT. Block: BlockOut Buffer (p/n orb348644). Exposure: 60 secs.

ELISA results of purified Goat Anti-Rabbit IgG F(c) Antibody (Min x Human serum proteins) tested against purified Rabbit IgG F(c) (Red Line), Rabbit IgG (Green Line), and Human IgG (Blue Line). Each well was coated in duplicate with 1.0 µg of antigen Rabbit IgG F(c) (orb346310), Rabbit IgG (orb2652747), and Human IgG (orb346219). The starting dilution of antibody was 5 µg/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 Blocking buffer, Donkey Anti-Goat IgG HRP (p/n orb347032), and TMB substrate (p/n orb348651).

Western Blot of Anti-Rabbit IgG F(c) (GOAT) Antibody (p/n orb347661). 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 of IgG, F(ab), IgM and Serum, 100 ng of F(c). Block: orb348637 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG F(c) (GOAT) Antibody (p/n orb347661) 1:2000 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 Goat anti-Rabbit Fc antibody. Lane 1: Rabbit Fc (p/n orb346310). Load: 100 ng per lane. Primary antibody: Rabbit Fc antibody at 1:1000 for overnight at 4°C. Secondary antibody: DyLight™ 649 goat secondary antibody at 1:20000 for 30 min at RT. Block: orb348637 for 30 min at RT. Predicted/Observed size: 28 kDa, 28 kDa for Rabbit Fc. Other band(s): Rabbit Fc splice variants and isoforms.