Invitrogen™ Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP, 1mL

The Invitrogen™ Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Catalog #31464) is a highly specific and affinity-purified secondary antibody designed for detecting rabbit primary antibodies in various immunoassays. This antibody is conjugated to horseradish peroxidase (HRP), enabling chemiluminescent, colorimetric, and fluorescent detection in Western blot (WB), immunohistochemistry (IHC), and immunocytochemistry (ICC/IF) applications.

This secondary antibody is cross-adsorbed to remove unwanted cross-reactivity with human, goat, mouse, and sheep serum proteins, ensuring high specificity and minimal background interference. It reacts specifically with whole rabbit IgG molecules and light chains of other rabbit immunoglobulins, making it a reliable and versatile reagent for signal amplification.

Applications & Recommended Dilutions

This antibody is optimized for multiple immunoassay applications, providing high sensitivity and minimal background noise:

1. Western Blot (WB):
   • Recommended dilution: 1:10,000 - 1:200,000
   • Detects rabbit primary antibodies bound to target proteins using HRP-based chemiluminescent substrates.
2. Immunohistochemistry (IHC):
   • Recommended dilution: 1:500 - 1:5,000
   • Used for detecting rabbit primary antibodies in tissue sections, providing highly specific staining with minimal background.
3. Immunocytochemistry / Immunofluorescence (ICC/IF):
   • Recommended dilution: 1:500 - 1:5,000
   • Enables visualization of rabbit primary antibodies in fixed or live cells with enzyme-based colorimetric detection.

Product Specifications

Key Features & Benefits

1. High Specificity & Cross-Adsorption:
   • Minimizes non-specific binding with cross-adsorption against human, goat, mouse, and sheep proteins.
   • Ensures clearer signals and reduced background noise in immunoassays.
2. HRP Conjugation for Sensitive Detection:
   • Compatible with chemiluminescent (ECL), colorimetric (DAB, TMB), and fluorometric HRP substrates.
   • Provides strong signal amplification for low-abundance targets.
3. Versatile Across Multiple Applications:
   • Validated for Western blot (WB), immunohistochemistry (IHC), and immunocytochemistry (ICC/IF).
   • Works in various immunoassay platforms with high sensitivity and reliability.
4. Lyophilized for Long-Term Stability:
   • Reconstitutes to 0.8 mg/mL in 1.0 mL distilled water.
   • Can be stored at 4°C for short-term use and at -20°C in 50% glycerol for long-term preservation.
5. Wide Dynamic Range in Western Blotting:
   • Effective at high dilutions (up to 1:200,000) for optimal signal-to-noise ratio.

Storage & Handling Instructions

• Before Reconstitution: Store at 4°C in its lyophilized form.
• After Reconstitution:
  • Store at 4°C for several weeks as an undiluted liquid.
  • If diluted, use within one day.
  • For long-term storage, add an equal volume of glycerol (final 50%) and store at -20°C.
• Clarity Check:
  • If the product is not completely clear after standing for 1-2 hours at room temperature, centrifuge it briefly.

Scientific Background: Anti-Rabbit Secondary Antibodies

Secondary antibodies play a crucial role in immunodetection assays by binding to primary antibodies and amplifying the signal. The Invitrogen™ Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP is designed to detect rabbit IgG (H+L) while ensuring minimal cross-reactivity with other species.

Why Cross-Adsorbed Secondary Antibodies?

• Minimizes species cross-reactivity for cleaner, more specific results.
• Prevents non-specific binding to endogenous immunoglobulins in tissue samples.
• Ideal for experiments requiring high specificity, such as co-localization studies and multiplex immunoassays.

HRP-Conjugated Secondary Antibodies in Immunodetection

• HRP (Horseradish Peroxidase) catalyzes substrate reactions to produce a detectable signal in Western blot, IHC, and ICC/IF.
• Enables highly sensitive detection with chemiluminescent, chromogenic, or fluorescent substrates.

Applications in Research

1. Western Blot Analysis (WB)
   • Detects rabbit primary antibodies bound to target proteins in tissue and cell lysates.
   • Provides high sensitivity for low-abundance proteins.
2. Immunohistochemistry (IHC)
   • Enables tissue-specific detection of target proteins using rabbit primary antibodies.
   • Useful in cancer research, histopathology, and biomarker studies.
3. Immunocytochemistry / Immunofluorescence (ICC/IF)
   • Facilitates cellular localization of proteins using enzyme-based detection.
   • Works well with fluorescent and colorimetric HRP substrates.
4. Signal Amplification in ELISA & Immunoprecipitation
   • Enhances sensitivity in sandwich and indirect ELISA assays.
   • Ensures robust detection of rabbit IgG antibodies in immunoprecipitation workflows.

Recommended Complementary Products

For optimal performance in immunoassays, consider the following Invitrogen secondary antibodies and reagents:

The Invitrogen™ Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Catalog #31464) is a high-performance, highly specific secondary antibody ideal for Western blot (WB), immunohistochemistry (IHC), and immunocytochemistry (ICC/IF). With HRP conjugation, cross-adsorption for minimal background, and broad application compatibility, this antibody is a powerful tool for immunodetection assays.