IP Lysis Buffer, IC-8100

Product Introduction

This product is a non-denaturing cell lysis buffer specifically formulated for immunoprecipitation (IP) and co-immunoprecipitation (Co-IP) applications. IP Lysis Buffer is composed of 25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, and 5% glycerol. The mild non-ionic detergent NP-40 effectively lyses cell membranes and solubilizes proteins while preserving native protein conformation and protein-protein interactions, making this buffer suitable for experiments requiring intact protein complexes. The formulation provides physiological ionic strength and pH for maintaining protein stability and antibody-antigen binding. The buffer is compatible with downstream applications including immunoprecipitation, co-immunoprecipitation, Western blot, and enzyme activity assays.

Product Features

1. Non-Denaturing Formulation: Mild NP-40 detergent preserves native protein conformation and protein-protein interactions.

2. Optimized for IP/Co-IP: Physiological ionic strength and pH support antibody-antigen binding and protein complex stability.

3. Ready-to-Use: Supplied as a ready-to-use solution requiring no additional preparation.

4. Broad Compatibility: Suitable for immunoprecipitation, co-immunoprecipitation, Western blot, and enzyme activity assays.

Specifications

Size: 100 mL / 250 mL

Formulation: 25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 5% glycerol

Application: Immunoprecipitation, co-immunoprecipitation, Western blot, enzyme activity assays

Storage and Stability

Storage Conditions: Store at 4¡ãC or -20¡ãC, protected from light. Avoid repeated freeze-thaw cycles.

Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed.

Protocol (For Reference Only)

Important: Pre-chill the buffer on ice before use. Add protease and phosphatase inhibitors freshly before cell lysis.

1. Buffer Preparation: Pre-chill the required volume of IP Lysis Buffer on ice. Add protease inhibitor cocktail (and phosphatase inhibitors if detecting phosphorylated proteins) immediately before use.

2. Cell Lysis: For adherent cells, wash twice with ice-cold PBS, aspirate residual liquid, and add an appropriate volume of ice-cold IP Lysis Buffer (typically 0.5-1 mL per 10⁷ cells). Incubate on ice for 15-30 minutes with occasional gentle rocking. Scrape cells and transfer the lysate to a pre-chilled microcentrifuge tube.

3. Clarification: Centrifuge at 12,000-14,000 x g for 10-15 minutes at 4¡ãC. Transfer the supernatant to a new pre-chilled tube.

4. Immunoprecipitation: Incubate the clarified lysate with the capture antibody and protein A/G beads according to standard IP/Co-IP protocols.

5. Downstream Analysis: Wash the beads with IP Lysis Buffer or a compatible wash buffer, elute the immunoprecipitated proteins, and analyze by SDS-PAGE and Western blot.

Precautions

1. Add protease and phosphatase inhibitors freshly before use.

2. Perform all steps at 4¡ãC or on ice to prevent protein degradation.

3. Avoid repeated freeze-thaw cycles; aliquot into single-use volumes.

4. For Western blot, add SDS-PAGE loading buffer and boil to denature before loading.

5. For research use only. Not for use in diagnostic or therapeutic procedures.

FAQ (Simplified)

Q1: What is the composition of this IP Lysis Buffer?

A1: It contains 25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, and 5% glycerol.

Q2: Is this buffer suitable for co-immunoprecipitation?

A2: Yes. The mild non-ionic detergent NP-40 preserves native protein conformation and protein-protein interactions, making it suitable for Co-IP.

Q3: How should I store this buffer?

A3: Store at 4¡ãC or -20¡ãC protected from light. Avoid repeated freeze-thaw cycles.

Q4: Do I need to add protease inhibitors?

A4: Yes. Add protease inhibitor cocktail (and phosphatase inhibitors if needed) freshly before cell lysis.

Disclaimer

1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.

2. Due to the variable nature of biological research, optimization of lysis conditions is recommended for specific cell types and protein targets.

3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages.

4. Wear appropriate protective clothing and gloves when handling this product.

Ordering Information

Catalog Number: IC-8100

Product Name: IP Lysis Buffer

Size: 100 mL / 250 mL

Suggested Retail Price:

100 mL: CNY ¥1550.00 / USD $155.00 / EUR €186.00 / JPY ¥27900.00

250 mL: CNY ¥2400.00 / USD $240.00 / EUR €288.00 / JPY ¥43200.00

Note: The specific price is subject to the transaction price in the offline contract. This document is provided for technical reference only and does not constitute any form of sales advertisement or invitation to offer. For product details and purchase inquiries, please contact local sales representatives.