CHAPS IMMUNOPRECIPITATION (IP) AND LYSIS BUFFER For Tissues and Cells

Chaps Lysis Buffer: Cell Lysis Buffer with CHAPS Detergent: Immunoprecipitation (IP) and Cell Lysis Buffer with CHAPS Detergent (IP Lysis Buffer)

Updated 5-2024

Protein extraction and immunoprecipitation for tissue and cell culture samples

CHAPS immunoprecipitation and lysis buffer with 0.5% CHAPS, HEPES, NaCl, 1X Solution. Preserves native protein conformation

Available Volumes

Volume	Cost
60 ml	$175
120 ml	$275

Available Formulations

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

*All pH's are offered at the same price.

The addition of calcium chloride to the CHAPS reagent preserves calcium dependent protein-protein interaction, whereas the addition of EDTA abrogates the interaction.

Highlights: CHAPS Buffer

Strong cell lysis and protein extraction while preserving protein conformation and protein-protein interaction for co-immunoprecipitation (Co-IP) and pull-down analysis.
Applicable with tissues (see product citation 1 below) as well as culture cells.
Effective as a stand alone reagent for cell lysis, protein extraction and immunoprecipitation with subsequent Western blot.
Contains a non-amine buffer (HEPES) enabling cross-linking with NHS-ester derivatives.
Ingredients: 0.5% CHAPS, HEPES, NaCl
Compatible with Bradford, Lowry and BCA protein assays.
60 ml volume: Suitable for two hundred 10 cm culture plates, or six hundred wells in 6-well dishes. 120 ml volume for four hundred 10 cm culture plates or 1200 wells in 6-well dishes.

IP and Lysis Protocol

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 summarized in the product manual. 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.

Outline of immunoprecipitation methodology with CHAPS Buffer (see details in protocol)

©Fivephoton Biochemicals 2010

Product Citations

1. Thakur, A., Park, K., Cullum, R. et al. (2024) HNF4A guides the MLL4 complex to establish and maintain H3K4me1 at gene regulatory elements. Commun Biol 7, 144. Link to article

2. Takata, Katsuyoshi, et al. (2022) "Tumor-associated antigen PRAME exhibits dualistic functions that are targetable in diffuse large B cell lymphoma." The Journal of Clinical Investigation 132.10 (2022). Link to article

3. Christopher R. Donnelly, et al. (2021). STING controls nociception via type I interferon signalling in sensory neurons. Nature volume 591, 275–280. Link to article

4. Joe Truong Nguyen, (2018). Mammalian EAK-7 activates alternative mTOR signaling to regulate cell proliferation and migration. Science Advances 09 May 2018: Vol. 4, no. 5, eaao5838. Link to article

5. Drum, Benjamin ML, et al. "Oxidative stress decreases microtubule growth and stability in ventricular myocytes." Journal of molecular and cellular cardiology 93 (2016): 32-43. Link to article

6. Villa‐Diaz, Luis G., et al. "Inhibition of Focal Adhesion Kinase Signaling by Integrin α6β1 Supports Human Pluripotent Stem Cell Self‐Renewal." Stem Cells (2016). Link to article

7. Kurland, D. B., Gerzanich, V., Karimy, J. K., Woo, S. K., Vennekens, R., Freichel, M., ... & Simard, J. M. (2016). The Sur1-Trpm4 channel regulates NOS2 transcription in TLR4-activated microglia. Journal of Neuroinflammation, 13(1). Link to article

8. Qian, X., Kim, J.K., Tong, W., Villa‐Diaz, L.G. and Krebsbach, P.H., 2015. DPPA5 Supports Pluripotency and Reprogramming by Regulating NANOG Turnover. STEM CELLS. Link to article

9. Zhang G, Brady J, Liang W-C, Wu Y, Henkemeyer M, Yan M. EphB4 forward signalling regulates lymphatic valve development. Nature Communications. 2015;6:6625. doi:10.1038/ncomms7625.  Link to article

10. Shuchong Pan et. al.  2014.  Cell Surface Protein Disulfide Isomerase  Regulates Natriuretic Peptide Generation of Cyclic Guanosine Monophosphate.  Plos One.  DOI: 10.1371/journal.pone.0112986.  Link to article

IP and Lysis Protocol

Accessory Products

Protease Inhibitor Cocktail (100X)

Phosphatase Inhibitor Cocktail (100X)

Sodium Orthovanadate (Preactivated, 200mM)

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

Storage: 4^oC on arrival.

Shipping: Ships at ambient temperature. International delivery is available.

kw:

IP Lysis Buffer, CHAPS Buffer, Immunoprecipitation, Protein extraction, Cell lysis, Protein, IP, Calcium binding protein, Co-IP, Pull down assay

Category	Download Link
Protocol Manual	click here
MSDS	click here

CHAPS IMMUNOPRECIPITATION (IP) AND LYSIS BUFFER For Tissues and Cells | Part No. CIB-1

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Transmembrane Protein Extraction Reagent (Part No. tmPER-50; Plasma Protein Isolation) TmPER-50 (100 plates) $180.00	TRANSMEMBRANE PROTEIN EXTRACTION REAGENT (tmPER-100; Membrane Protein Isolation) TmPER-100 (150 plates) $200.00	Transmembrane Protein Extraction Reagent (tmPER-200; Plasma Protein Isolation) TmPER-200 (300 plates) $260.00	SODIUM ORTHOVANADATE, ACTIVATED, 200mM SOLN \| Part ActVO-4 ActVO-4 Activated $145.00
RIPA Buffer (RIPA-50) RIPA-50 $82.00	PROTEASE INHIBITOR COCKTAIL BROAD SPECTRUM (100X; PI-1) PI-1 $115.95	PROTEASE INHIBITOR COCKTAIL w/ EDTA (100X, 1ml; PI-2) PI-2 $115.95	Tissue Protein Extraction Buffer (1X, 100 ml; Part No TEB-1) TEB-1 $125.00

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CHAPS IMMUNOPRECIPITATION (IP) AND LYSIS BUFFER For Tissues and Cells | Part No. CIB-1