Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody

Western Blot: Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody [NB120-6824] - Lane 1: Rabbit IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Peroxidase goat secondary antibody at 1:1,000 for 60 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Rabbit IgG. Other band(s): None. Image using the HRP form of this antibody.

Western Blot: Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody [NB120-6824] - Lane 1: Rabbit IgG. Lane 2: None. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Fluorescein goat secondary antibody at 1:1,000 for 60 min at RT. Blocking: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 25 & 55 kDa, 25 & 55 kDa for Rabbit IgG. Other band(s): None.

SDS-Page: Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody [NB120-6824] - SDS PAGE of Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody [Unconjugated].Lane 1: 1x Reduced, Fab Anti-RABBIT IgG Antibody.Lane 2: Molecular Weight Ladder.Lane 3: 1x Non-Reduced, Fab Anti-RABBIT IgG Antibody.4-20% Lonza SDS-PAGE; Coomassie Stained.

• Reactivity: Rb
• Applications: WB, ELISA, IHC, PAGE
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated

Goat F(ab) anti-Rabbit IgG (H+L) Secondary Antibody 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, papain digestion and chromatographic separation. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum
  
  Store vial at 4C prior to opening. This product is stable at 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: Rabbit IgG whole molecule
• Specificity: No reaction was observed against anti-Papain or anti-Goat IgG F(c).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:4000 - 1:20000
  • Immunohistochemistry 1:1000 - 1:5000
  • SDS-Page
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product has been tested by dot blot and SDS-PAGE and is suitable for highly specific immunological methods requiring extremely low background levels, absence of F(c) mediated binding, lot-to-lot consistency, high titer and specificity.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Background

Antibodies, also known as immunoglobulins (Igs) are critical for immunity and are grouped into five primary classes: IgG, IgM, IgA, IgD, and IgE. The most abundant antibody isotype is immunoglobulin G (IgG) with concentrations ranging from 7.5-22 mg/ml in human serum and has a molecular weight of 150 kDa. The major effector functions of IgG include neutralization, opsonization, complement fixation and antibody dependent cell-mediated cytotoxicity (ADCC). This monomeric immunoglobulin, expressed on the surface of mature B cells, is often depicted as a Y-shape and comprised of 2 heavy chains and 2 light chains linked by disulfide bonds. The heavy chain is type gamma including subtypes gamma 1, gamma 2, gamma 3, and gamma 4 while the light chain is either a kappa or lambda chain. An IgG molecule has two antigen binding sites, each consisting of a heavy and light chain N-terminal variable domain. When combined with the constant heavy chain 1 (Ch1) and the constant light chain domains, it forms the fragment antigen-binding (Fab) region (2 per antibody). The remaining domains (Ch2-Ch4) of both heavy chains make up the Fc region and contain a site for covalently linking an enzymatic or fluorochrome probe, such as HRP or Janelia Fluor 549, for target detection and visualization (1,2,3).

The 4 IgG subclasses, sharing 95% amino acid identity, include IgG1, IgG2, IgG3, and IgG4 for humans and IgG1, IgG2a, IgG2b, and IgG3 for mice. The relative abundance of each human subclass is 60% for IgG1, 32% for IgG2, 4% for IgG3, and 4% for IgG4. In an IgG deficiency, there may be a shortage of one or more subclasses (4).

References

1. Painter RH. (1998) Encyclopedia of Immunology (Second Edition). Elsevier. 1208-1211

2. Chapter 9 - Antibodies. (2012) Immunology for Pharmacy. Mosby 70-78

3. Schroeder H, Cavacini, L. (2010) Structure and Function of Immunoglobulins. J Allergy Clin Immunol. 125(2 0 2): S41-S52. PMID: 20176268

4. Vidarsson G, Dekkers G, Rispens T. (2014) IgG subclasses and allotypes: from structure to effector functions. Front Immunol. 5:520. PMID: 25368619

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.

FAQs for IgG (H+L) Antibody (NB120-6824). (Showing 1 - 1 of 1 FAQ).

1. May we ask if NB120-6824 is the antibody against Fab2 region of Rabbit IgG? We are confused because there is no further description about its specificity for Fab2, and its immunogen is "Rabbit IgG whole molecule." Would you please help confirm?
   • This is a Fab fragment of an antibody directed against whole Rabbit IgG. Yes this antibody is the Fab fragment of goat IgG and recognizes whole Rabbit IgG.