CHAPS Lysis and Immunoprecipitation Buffer | FIVEphoton Biochemicals | CHAPS Buffer 1X

CHAPS Lysis and Immunoprecipitation (IP) Buffer

CHAPS cell lysis and immunoprecipitation buffer with 0.5% CHAPS, HEPES, 1X Solution

(Formulations available. CHAPS buffer, pH 8.0; CHAPS buffer, pH 7.4; CHAPS buffer, pH 6.0; optional addition of 1mM EDTA; optional addition of 1mM CaCl₂).

Highlights, CHAPS Buffer

Strong cell lysis and protein extraction capacity while preserving protein conformation and protein-protein interaction for co-immunoprecipitation (Co-IP) analysis.
Effective as a stand alone reagent for cell lysis and protein extraction.
Contains a non-amine buffer (HEPES) enabling cross-linking with NHS-ester derivatives.
Minimal interference with Bradford and Lowry protein assays.
60 ml volume: Suitable for two hundred 10-cm culture plates, or six hundred wells in 6-well dishes. 120 ml volume also available.

View Protocol

Properties of CHAPS Lysis and Immunoprecipitation Buffer

CHAPS is a zwitterionic detergent with strong membrane dissolution capacity while maintaining protein conformation and protein-protein interaction. The CHAPS reagent provides an effective stand alone buffer for cell lysis and protein extraction as well as a reagent suitable for downstream immunoprecipitation-pull down approaches.

Optional Formulations, Ingredients and Volumes in the CHAPS Buffer to Investigate Calcium Dependent Interactions

Cost

Volume	Cost
60 ml	$68.50
120 ml	$120.50

CHAPS Buffer pH Options*

Part No	pH
Part No.CIB-1-8.0	pH 8.0
Part No. CIB-1-7.4	pH 7.4
Part No CIB-1-6.0	pH 6.0

*All pH's are offered at the same price. See Available Options menu to order.

CaCl₂ and EDTA Options*

Optional additon of 1 mM EDTA

Optional addition of 1 mM CaCl₂

_{*Addition of CaCl2 or EDTA does not affect price. See "Available Options" menu to order.}

The addition of calcium chloride to the CHAPS reagent preserves calcium dependent protein-protein interaction, whereas separately, the addition of EDTA abrogates the interaction. The CHAPS immunoprecipitation reagent is offered with or without 1 mM calcium chloride and/or 1mM EDTA to investigate the roles of calcium in protein-protein interactions. Use the Available Options menu above to select the calcium chloride or EDTA formulations in the CHAPS Reagent.

(*Note: The CHAPS Buffer with formulations containing calcium chloride, or EDTA, have been employed to characterize calcium dependent chaperone protein interactions, such as with calnexin or calreticulin (see references 2-6 below).

Representative Data with CHAPS Immunoprecipitation and Cell Lysis Reagent (pH 8.0)

Figure 1. Co-immunoprecipitation (Co-IP) of a trafficking protein receptor (COPII) with a cation channel (TRPC-6). HEK293 cells were co-transfected with plasmid DNA encoding TPRC-6 and a HA epitope tagged component of COPII (sec24). Cell lysis and immunoprecipitation were performed in CHAPS Buffer as indicated in the product protocol. Western blotting (WB) was performed with the antibodies indicated in the figure.

IP: Anti-HA COPII IP: Anti-HA COPII

WB: Anti-TRPC6 WB: COPII

Figure 2. Co-immunoprecipitation (Co-IP) of a calcium binding protein with a multisubunit target protein 1 employing the FIVEphoton CHAPS immunoprecipitation reagent. A larger amount of target protein 1 associates with the calcium binding protein when expressed alone, as compared to co-expression (and presumed assembly) with protein 2. Protein 1 reveals higher molecular weight forms due to post-translational modifications.

Formulations Available. All CHAPS Buffer formulations are provided 1X

Use "Available Options" menu above to select the following in the CHAPS IP Reagent. On Purchase Orders, please note pH of CHAPS buffer and whether EDTA or calcium chloride is requested.

1. pH 8.0

2. pH 7.4

3. pH 8.0

4. No EDTA or CaCl₂

5 With 1mM EDTA

6. With 1mM CaCl₂ (no EDTA)

Outline of Immunoprecipitation Methodology with CHAPS Buffer (see protocol for details)

References on CHAPS Immunoprecipitation and Cell Lysis

Darby PJ, Kwan CY, Daniel EE. 2000. Caveolae from canine airway smooth muscle contain the necessary components for a role in Ca(2+) handling. Am J Physiol Lung Cell Mol Physiol 279: L1226-35.
Farmery MR, Allen S, Allen AJ, Bulleid NJ. 2000. The role of ERp57 in disulfide bond formation during the assembly of major histocompatibility complex class I in a synchronized semipermeabilized cell translation system. J Biol Chem 275: 14933-8.
Ora A, Helenius A. 1995. Calnexin fails to associate with substrate proteins in glucosidase-deficient cell lines. J Biol Chem 270: 26060-2.
Pieren M, Gali C, et. al. 2005. The use of calnexin and calreticulin in cellular and viral glycoproteins. J Biol Chem 280: 28265-71.
Zhu X, Peng J, Chen D, Liu X, Ye L, et al. 2005. Calnexin and ERp57 facilitate the assembly of the neonatal Fc. Journal of Immunology 175: 967-76.

Safety: Irritant. Avoid skin and eye contact and ingestion.

Storage: 4^oC.

Shipping: Domestic Fedex 2-day (flat rate $18) or Fedex overnight delivery (flat rate $25). International delivery is also available. Please inquire for international shipping rates.

Category	Download Link
Protocol Manual	click here
MSDS	click here

CHAPS Lysis and Immunoprecipitation Buffer | FIVEphoton Biochemicals | CHAPS Buffer 1X | CIB-1

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CHAPS Lysis and Immunoprecipitation Buffer | FIVEphoton Biochemicals | CHAPS Buffer 1X | CIB-1