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Specifications

Zebrafish embryos were collected 48 hours post fertilization (48 hpf), homogenized in modified RIPA buffer and cleared by centrifugation. Prior to lysis, embryos were treated to remove majority of the embryonic yolk. Total protein concentration of the embryo lysate was determined using BCA protein assay (BSA was used as a standard). Lysate was mixed with 2X Laemmli sample buffer containing reducing reagent and heated for 5 minutes at 90°C. E2256-20A is supplied at 2mg/ml total protein concentration in 1x laemmli sample buffer.
Catalog #	E2256-20A
Applications	Suitable for use in Western Blot and SDS-PAGE electrophoresis. Other applications not tested.
Recommended Dilution	Western Blot: 15-20ug of total protein per lane
	Optimal dilutions to be determined by the researcher.
Storage and Stability	Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Source	Wild-type zebrafish embryos
Purity	Lysate
Concentration	~2mg/ml
Form	Supplied as a liquid in RIPA buffer and 1x Laemmli sample buffer. (RIPA buffer: 150mM sodium chloride, 50mM Tris-Cl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 1mM EDTA, 1mM PMSF, 5ug/ml Pepstatin A, 2ug/ml Leupeptin, 1mM sodium orthovanadate, 1mM sodium fluoride).
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.