I use your SNAP-25 protein routinely in my experiments using a frozen stock that I keep at -80C. It works fine. However, for my experiments I cleave the SNAP-25 (lop off the last 10 aa or so) and use this protein for my ELISA. I cleave the SNAP-25 by incubating it for 2 hours with the 50KD Enzymatic light chain from Botulinium toxin A. The fresh cleaved SNAP-25 works fine in my ELISA. When I take 0.5 mg/ml of the cleaved SNAP-25 (also containing 0.25 mg/ml toxin A) and freeze it at -80C in 25 mM Tris-HCL, 1 mM DTT, 2 mM EDTA, and 10% glycerol (your formulation for SNAP-25) I lose 70% of my activity in 3 days. Do you have any tips on how to store this cleaved SNAP-25? Preparing the product fresh every time is not practical!

Name: Recombinant Human SNAP25 Protein
Brand: Novus Biologicals
SKU: NBC1-18344
Price: 409 USD

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I imagine that the loss of activity comes from storing the protein in the presence of Botulinum toxin A and not from freeze thawing. Have you tested that activity remains after a round of freezing and thawing in the absence of the toxin? In this situation, I think you need to remove the Botulinum toxin since even at -80C, the toxin could be continuing to cleave and destroy the protein. The toxin you said is 50kDa, and the protein is 25kDa. Could you remove the toxin by size exclusion chromatography? Additionally, I found this paper, Procedures for the Inactivation and Safe Containment of Toxins (http://www.ouhsc.edu/ehso/toxinproc.pdf) that suggests that you can inactivate the toxin by heating or by adding sodium hypochorite and/or sodium hydroxide. I know this protein can be used for Western blot, so it should be able to survive boiling but 200 degrees Celsius may be too much but could be worth trying on an aliquot? Size exclusion chromatography, bleaching and heating are our three suggestions at the moment. If none of these work, then you may have to continue to prepare it fresh.

At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Reactivity	HuSpecies Glossary
Applications	PAGE
Concentration	1 mg/ml

Description	An un-tagged recombinant protein corresponding to the amino acids1-206 of Human SNAP25 Source: E.coli Amino Acid Sequence: MAEDADMRNE LEEMQRRADQ LADESLESTR RMLQLVEESK DAGIRTLVML DEQGEQLERI EEGMDQINKD MKEAEKNLTD LGKFCGLCVC PCNKLKSSDA YKKAWGNNQD GVVASQPARV VDEREQMAIS GGFIRRVTND ARENEMDENL EQVSGIIGNL RHMALDMGNE IDTQNRQIDR IMEKADSNKT RIDEANQRAT KMLGSG
Source	E. coli
Protein/Peptide Type	Recombinant Protein
Gene	SNAP25
Purity	>90%, by SDS-PAGE

Dilutions	SDS-Page
Theoretical MW	23.3 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Storage	Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer	25mM Tris-HCl buffer (pH 7.5), 1 mM DTT, 10% glycerol
Preservative	No Preservative
Concentration	1 mg/ml
Purity	>90%, by SDS-PAGE

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