Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed)

Western Blot: Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed) [NBP2-60663] - Western Blot of Goat anti-Rabbit IgG Fc Secondary antibody (Pre-adsorbed). Lane 1: Rabbit Fc. Load: 100 ng per lane.Primary antibody: Rabbit Fc antibody at 1:1,000 for overnight at 4C.Secondary antibody: DyLight(TM) 649 goat secondary antibody at 1:20,000 for 30 min at RT.Block for 30 min at RT.Predicted/Observed size: 28 kDa, 28 kDa for Rabbit Fc. Other band(s): Rabbit Fc splice variants and isoforms.

Western Blot: Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed) [NBP2-60663] - Lane 1: Rabbit IgG whole molecule. Lane 2: Rabbit IgG F(ab) Fragment. Lane 3: Rabbit IgG F(c) Fragment. Lane 4: Rabbit IgM Whole Molecule. 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: incubated with blocking buffer for 30 min at RT. Primary Antibody: Anti-Rabbit IgG F(c) (GOAT) Antibody 1:2,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody 1:40,000 in incubated with blocking buffer 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. Image using the Biotin form of this antibody.

Dot Blot: Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed) [NBP2-60663]

ELISA: Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed) [NBP2-60663] - ELISA results of purified Goat anti-Rabbit IgG Fc Secondary antibody (Pre-adsorbed) tested against purified RabbitIgG Fc (Red Line), Rabbit IgG (Green Line), and Human IgG (Blue Line). Each well was coated in duplicate with 1.0 ug of antigen RabbitIgG Fc, Rabbit IgG, and Human IgG. The starting dilution of antibody was 5ug/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, and TMB substrate.

• Reactivity: Rb
• Applications: WB, DB, ELISA, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: Pre-adsorbed

Goat anti-Rabbit IgG Fc Secondary Antibody (Pre-adsorbed) Summary

• Description: 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
  
  Store vial at 4C prior to opening. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.
• Immunogen: Goat anti-Rabbit IgG Fc fragment Secondary Antibody (Pre-adsorbed) was produced by repeated immunization with rabbit IgG F(c) fragment in goat.
• Specificity: This antibody was pre-adsorbed against Human Serum Proteins.
  
  No reaction was observed against Rabbit IgG F(ab) or Human Serum Proteins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA 1:20000 - 1:100000
  • Immunohistochemistry 1:1000 - 1:5000
  • Immunohistochemistry-Paraffin 1:50 - 1:200
  • Western Blot 1:1000-1:1:10000
• 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Secondary Antibodies are guaranteed for 1 year from date of receipt.