Type
Recombinant protein
Description
Total 201 AA. MW: 23.4 kDa (calculated). N-terminal His-tag, 10 extra AA (highlighted). The AA sequence is identical to UniProtKB/Swiss-Prot entry P62760.
Amino Acid Sequence
MKHHHHHHASMGKQNSKLAPEVMEDLVKSTEFNEHELKQWYKGFLKDCPSGRLNLEEFQQLYVKFFPYGDASKFAQHAFRTFDKNGDGTIDFREFICALSITSRGSFEQKLNWAFNMYDLDGDGKITRVEMLEIIEAIYKMVGTVIMMKMNEDGLTPEQRVDKIFSKMDKNKDDQITLDEFKEAAKSDPSIVLLLQCDIQK
Source
E. coli
Purity
Purity as determined by densitometric image analysis: > 95%
SDS-PAGE Gel
12% SDS-PAGE separation of Human VILIP
1. M.W. marker – 14, 21, 31, 45, 66, 97 kDa
2. reduced and heated sample, 5μg/lane
3. non-reduced and non-heated sample, 5μg/lane
Endotoxin
< 0.1 EU/μg
Formulation
Filtered (0.4 μm) and lyophilized from 0.5 mg/ml in 20mM Tris buffer, 50mM NaCl, pH 7.5
Reconstitution
Add deionized water to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. Product is not sterile! Please filter the product by an appropriate sterile filter before using it in the cell culture.
Applications
Western blotting
Shipping
At ambient temperature. Upon receipt, store the product at the temperature recommended below.
Storage/Expiration
Store the lyophilized protein at –80 °C. Lyophilized protein remains stable until the expiry date when stored at –80 °C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80 °C for long term storage. Reconstituted protein can be stored at 4 °C for a week.
Quality Control Test
BCA to determine quantity of the protein.
SDS PAGE to determine purity of the protein.
LAL to determine quantity of endotoxin.
Note
This product is intended for research use only.
Research topic
Neural tissue markers, Oncology
Summary
Visinin like protein 1 (VILIP-1, VLP-1 or VSNL-1) is a cytoplasmic protein of low molecular weight (approximately 22 kDa) consisting of 191 amino acid residues. It belongs to the visinin/recoverin subfamily of neuronal calcium sensor proteins involved in calcium-dependent signal transduction mechanisms in neurons. It is found primarily in the brain, in nerve cells, but it also has a peripheral distribution in liver, lung, kidney, spleen, pancreas and colon. When localized at the membrane, it modulates various cellular signal transduction pathways, including cyclic adenosine monophosphate (cAMP)- and cyclic guanosine monophosphate (cGMP)-signaling in neural cells, human embryonic kidney cells, the pancreatic β cell line MIN6, and various skin tumor cell lines. It contains four internal repeats of 36–38 amino-acids, each containing a potential EF-hand domain. Two of the four EF-hand Ca2+-binding motifs of VILIP-1 are able to bind either Ca2+ or Mg2+in a non-cooperative manner. Binding of Ca2+ leads to specific conformational changes in the protein and this may regulate the interaction of VILIP with intracellular target molecules. VILIP-1 has been identified as a potential biomarker for brain injury and several neurodegenerative diseases. VILIP-1-expressing cells appear to be vulnerable to neurotoxic insults. As a result, the protein is released into the cerebrospinal fluid (CSF), and can be used as a biomarker for stroke and Alzheimer’s disease. The intracellular protein was detected in cerebrospinal fluid (CSF) of a rat model of stroke and in plasma of patients after stroke. VILIP-1 was detected in 44% of subjects with stroke, in samples taken 24 hours after onset of stroke, and in 8% of controls with no stroke. In Alzheimer’s disease, CSF levels of VILIP-1 were significantly higher than in healthy individuals. Post mortem studies in the hippocampus of schizophrenia patients revealed increased expression of VILIP-1 in interneurons, while its expression in pyramidal neurons was downregulated. Expressions of VILIP-1 were also found in different types of cancer and in pancreatic α- and β-cells, being involved in the regulation of insulin secretion and insulin gene expression.
