PicoTect Western Blot Chemiluminescent Substrate Kit

Product Discontinued

PicoTect Western Blot Chemiluminescent Substrate Kit Summary

• Description: ADVANTAGES
  
• High intensity signal- twice as intense as other luminol-based systems
• Picogram sensitivity - highly sensitive for the rapid development of a wide range of protein levels
• Immunogen: ADVANTAGES
  High intensity signal- twice as intense as other luminol-based systems Picogram sensitivity - highly sensitive for the rapid development of a wide range of protein levels Excellent stability - 8-hour working solution stability
• Kit Type: Kit

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: For best results, it is ESSENTIAL to optimize all components of the system including sample amount, primary and secondary antibody concentration, and the choice of membrane and blocking reagents. PicoTectTM Western Blot Chemiluminescent Substrate is extremely sensitive, requiring less sample and primary and secondary antibodies than other commercially available substrates, usually by a factor of at least 10-20. Primary antibodies purchased from IMGENEX contain indications on the technical data sheet for use in Western Blot which are optimized for use with this substrate system. Blocking buffer formulations may need to be empirical tested to determine the appropriate conditions for each Western blot system. Determining the proper blocking buffer can help increase sensitivity and prevent nonspecific signal caused by cross-reactivity between the antibody and the blocking reagent. Furthermore, when switching from one substrate to another, a diminished signal or increased background sometimes results when the blocking buffer was not optimal for the new system. At IMGENEX we use 5% Carnation nonfat dry milk in TBST (25 mM Tris-Cl, pH 8.0; 125 mM NaCl; 0.1% Tween 20). Avoid using milk as a blocking reagent when using avidin/biotin systems because milk contains variable amounts of endogenous biotin. The PicoTect Substrate Working Solution is stable for 8 h at RT. Exposure to the sun or any other intense light can harm the Working Solution. For best results keep the Working Solution in an amber bottle and avoid prolonged exposure to any intense light. Short-term exposure to typical laboratory lighting will not harm the Working Solution. Use a sufficient volume of all buffers and solutions to cover blot and ensure that it never becomes dry. Large blocking and wash buffer volumes may result in reduction of specific signal. For optimal results, use a shaking platform during incubation steps. Do not use sodium azide as a preservative for buffers. Sodium azide is an inhibitor of HRP and could interfere with this system. Do not handle membrane with bare hands. Always wear gloves or use clean forceps. All equipment must be clean and free of foreign material. Metallic devices (e.g., scissors) must have no visible signs of rust. Rust may cause speckling and/or high background

Packaging, Storage & Formulations

• Storage: Storage is content dependent.

Notes

ADDITIONAL ITEMS REQUIRED (NOT INCLUDED IN THE KIT)
Immobilon P membranes (Millipore Corporation, MA). Nitrocellulose membrane or other completed western blot membrane types can be used; however, optimization may be required. Wash Buffer: TBST (25 mM Tris-Cl, pH 8; 125 mM NaCl; 0.1% Tween 20). Dilution Buffer: 1% Carnation nonfat dry milk in TBST. Blocking Reagent: 5% Carnation nonfat dry milk in TBST. Primary Antibody: Choose an antibody that is specific to the target protein(s) and prepare according to the manufactures specification. HRP-conjugated Secondary Antibody (azide-free): Choose an HRPconjugate that specifically binds to the primary antibody. The optimal dilution to use varies depending on the HRP conjugate and the amount of antigen on the membrane. Film cassette, developing and fixing reagents: For processing autoradiographic film. Rotary platform shaker: For agitation of membrane during incubations

Alternate Names for PicoTect Western Blot Chemiluminescent Substrate Kit

• Chemiluminescent Substrate
• PicoTect Western Blot
• Western Blot Substrate

Background

PicoTect Western Blot Chemiluminescent Substrate is a highly sensitive enhanced substrate for detecting horseradish peroxidase (HRP) on immunoblots. This substrate's extremely intense signal output enables detection of picogram amounts of antigen. The sensitivity, intensity and duration of the signal allows for easy detection of HRP using photographic or other imaging methods. Blots can also be repeatedly exposed to film to obtain optimal results or stripped of the immunodetection reagents and reprobed.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for PicoTect Western Blot Chemiluminescent Substrate Kit (NBP2-29912)(2)

Publications using NBP2-29912

• You MK, Kim HJ, Kook JH, Kim HA. St. John's Wort Regulates Proliferation and Apoptosis in MCF-7 Human Breast Cancer Cells by Inhibiting AMPK/mTOR and Activating the Mitochondrial Pathway. Int J Mol Sci. 2018-03-23 [PMID: 29570671] (WB)
• Rhyu J, Kim MS, You MK et al. Pear pomace water extract inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Nutr Res Pract 2014-02-01 [PMID: 24611103] (WB, Mouse)
  
  Details:
  3T3-L1, Figs 3, 4