Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed)

Western Blot: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - Lane 1: Molecular Weigh Ladder. Lane 2: Guinea Pig IgG. Lane 3: Bovine Serum. Lane 4: Chicken Serum. Lane 5: Goat Serum. Lane 6: Hamster Serum. Lane 7: Horse Serum. Lane 8: Human Serum. Lane 9: Mouse Serum. Lane 10: Rabbit Serum. Lane 11: Rat Serum. Lane 12: Sheep Serum. Loading: 50ng IgG, Serum at 1:8 dilution. Incubated in blocking buffer for 30 min at RT. Secondary Antibody: NB120-7139. Image from the HRP version of this antibody.

Immunocytochemistry/Immunofluorescence: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - Immunocytochemistry/Immunofluorescence: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody [DyLight 488] (Pre-absorbed) [NBP1-72854]

Dot Blot: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - Load: Lane 1 - 200 ng Lane 2 - 66.67 ng Lane 3 - 22.22 ng Lane 4 - 7.41 ng Lane 5 - 2.47 ng. Primary antibody: none. Secondary antibody: Goat anti-Guinea Pig IgG Antibody Alkaline Phosphatase Conjugated at 1:1,000 for 60 min at RT. Blocked with blocking buffer for 60 min at RT. Visualized using Alkaline Phosphatase Substrate for 30 seconds at RT. Image from the Alkaline Phosphatase version of this antibody.

Dot Blot: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - Dots are Guinea Pig IgG: (1) 100ng, (2) 33.3ng, (3) 11.1ng, (4) 3.70ng, (5) 1.23ng. Primary Antibody: none. Secondary Antibody: Goat Anti-Guinea Pig IgG Antibody FITC at 1ug/mL in blocking buffer for 1hr at RT. Imaged with BioRad ChemiDoc, Fluorescein filter. Image from the FITC version of this antibody.

ELISA: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - Using the DyLight 405 format of this antibody.

ELISA: Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) [NB120-7134] - ELISA Results of Goat anti-Guinea Porcine IgG (H+L) Secondary antibody (Pre-adsorbed) tested against purified Guinea Porcine MX10. Each well was coated in duplicate with 1.0 ug of Guinea Porcine IgG. The working dilution is 1:51,000. 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 HRP Conjugation Stabilizer, Donkey Anti-Goat IgG HRP conjugated, and TMB substrate.

• Reactivity: Gp
• Applications: WB, DB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: Pre-adsorbed

Goat anti-Guinea Pig IgG (H+L) Secondary Antibody (Pre-adsorbed) Summary

• Description: This product was prepared from monospecific antiserum by immunoaffinity chromatography using Guinea Pig 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, Guinea Pig IgG and Guinea Pig 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: Guinea Pig IgG whole molecule
• Specificity: This antibody was pre-adsorbed against Bovine, Chicken, Goat, Hamster, Horse, Human, Mouse, Rabbit, Rat, and Sheep Serum Proteins.
  
  No reaction was observed against Bovine, Chicken, Goat, Hamster, Horse, Human, Mouse, Rabbit, Rat and Sheep Serum Proteins. Minimal cross reactivity was observed against the species listed above.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA 1:50000
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:1000 - 1:5000
  • Western Blot 1:2000 - 1:10000
• Application Notes: This product is suitable for ELISA, western blot, and immunohistochemistry, as well as other assays requiring lot-to-lot consistency.

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

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.

Publications for IgG (H+L) Antibody (NB120-7134)(1)

Publication using NB120-7134

• Lopez-Murcia FJ, Reim K, Jahn O et al. Acute Complexin Knockout Abates Spontaneous and Evoked Transmitter Release Cell Rep 2019-03-07 [PMID: 30840877] (ICC/IF)