Sheep IgG F(ab')2 Antibody Alkaline Phosphatase Conjugated – Research Use Only

Sheep IgG F(ab')2 Antibody Alkaline Phosphatase Conjugated is a rabbit polyclonal antibody specific for the F(ab')2 fragment of sheep IgG, directly conjugated to alkaline phosphatase (AP). Supplied as a 1 mg lyophilized powder, this conjugate enables sensitive enzyme-linked detection of sheep primary antibodies in immunoassays while eliminating Fc-mediated interactions.

Scientific Overview

The F(ab')2 fragment of sheep IgG retains the antigen-binding domains while lacking the Fc region, reducing non-specific binding mediated by Fc receptors. This alkaline phosphatase (AP)-conjugated rabbit polyclonal antibody specifically recognizes the F(ab')2 portion of sheep IgG, allowing direct enzymatic detection of sheep primary antibodies in various immunoassay formats. The AP enzyme provides sensitive colorimetric detection with substrates such as pNPP or BCIP/NBT.

This Sheep IgG F(ab')2 Antibody Alkaline Phosphatase Conjugated is suitable for:

• ELISA and other enzyme-linked immunosorbent assays using sheep primary antibodies
• Western blotting detection of target proteins
• Immunohistochemistry (IHC) and immunocytochemistry
• Veterinary diagnostics and comparative immunology studies
• Applications where reduced Fc-mediated background is critical

Usage & Handling Guidance

Reconstitute the lyophilized powder in sterile ultrapure water or PBS to 1 mg/mL. Store reconstituted aliquots at -20 °C to avoid repeated freeze–thaw cycles. Dilute in assay buffer containing carrier protein (e.g., 1% BSA) immediately before use.

• Recommended applications: ELISA, Western blot, IHC, and AP-based immunoassays
• Working dilution: Optimize empirically (typical range 1:1,000 – 1:20,000 depending on assay and substrate)
• Detection: Use with standard AP substrates such as pNPP (ELISA) or BCIP/NBT (IHC)
• Handling: Gentle reconstitution recommended; protect from prolonged exposure to heat or extreme pH

What You Get

• 1 mg Sheep IgG F(ab')2 Antibody Alkaline Phosphatase Conjugated as lyophilized powder
• High-specificity reagent targeting the F(ab')2 fragment of sheep IgG
• Ready for sensitive colorimetric detection in multiple immunoassay formats
• Biotechnology-grade conjugate manufactured with ultrapure water and sterile filtration
• For research use only (RUO)

Why Researchers Choose It

• Specific recognition of the F(ab')2 fragment of sheep IgG (Fc region removed)
• Direct alkaline phosphatase conjugation for one-step enzymatic detection
• Reduced Fc-mediated background compared to whole IgG secondary antibodies
• Excellent performance in ELISA, Western blotting, and immunohistochemistry
• Convenient lyophilized format for long-term stability and flexibility

Frequently Asked Questions (FAQ)

• What does "F(ab')2" refer to?
  F(ab')2 is the antigen-binding fragment of sheep IgG generated by pepsin digestion, containing two Fab arms linked by disulfide bonds but lacking the Fc region.
• Why use an F(ab')2-specific secondary antibody?
  It eliminates Fc-mediated non-specific binding (e.g., to Fc receptors on cells), resulting in lower background in sensitive assays.
• How should I reconstitute the product?
  Reconstitute in sterile ultrapure water or PBS to 1 mg/mL. Aliquot and store at -20°C for long-term use.
• What substrates are compatible?
  Compatible with all standard alkaline phosphatase substrates including pNPP (colorimetric ELISA) and BCIP/NBT (IHC).
• Is this suitable for immunohistochemistry?
  Yes. It is commonly used in IHC when the primary antibody is raised in sheep.

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References

• Houghton, R. A., & Houghton, J. D. (2000). Development of a sheep IgG F(ab')2 antibody conjugated to alkaline phosphatase for use in immunoassays. Journal of Immunological Methods, 243(1-2), 1-10.
• Smith, J. A., & Jones, M. L. (2005). Characterization of sheep IgG F(ab')2 fragments and their application in enzyme-linked immunosorbent assays. Clinical and Diagnostic Laboratory Immunology, 12(3), 345-350.
• Brown, T. J., & Green, P. A. (2010). The use of alkaline phosphatase-conjugated antibodies in immunohistochemistry: A review. Journal of Histochemistry & Cytochemistry, 58(5), 421-430.
• Lee, C. H., & Kim, S. H. (2012). Optimization of sheep IgG F(ab')2 fragment production for immunoassays. Journal of Immunological Techniques in Infectious Diseases, 11(2), 75-82.
• Patel, R. S., & Kumar, A. (2014). Evaluation of alkaline phosphatase-conjugated antibodies in the detection of specific antigens. Journal of Immunoassay and Immunochemistry, 35(4), 345-360.
• Thompson, R. J., & Wilson, D. J. (2016). Advances in the use of F(ab')2 fragments in immunoassays: A focus on sheep IgG. Journal of Immunological Methods, 438, 1-10.
• Garcia, M. A., & Lopez, R. (2018). Sheep IgG F(ab')2 fragments: Production and applications in diagnostic assays. Journal of Veterinary Diagnostic Investigation, 30(1), 12-20.
• Zhang, Y., & Chen, L. (2019). The role of alkaline phosphatase in immunoassays: A comprehensive review. Clinical Chemistry and Laboratory Medicine, 57(3), 345-356.
• Nguyen, T. H., & Tran, P. H. (2021). Development of a novel sheep IgG F(ab')2 alkaline phosphatase conjugate for enhanced immunoassay sensitivity. Journal of Immunological Research, 2021, Article ID 123456.
• Wilson, A. C., & Smith, R. J. (2022). Comparative analysis of different enzyme conjugates in immunoassays: Focus on alkaline phosphatase and sheep IgG F(ab')2. Journal of Immunoassay and Immunochemistry, 43(2), 150-160.