Liver Expressed Antimicrobial Peptide 2 (LEAP2) Recombinant, Human

Recombinant protein corresponding to aa23-76 with two N-Terminal Tags, His-Tag and GST-Tag from human LEAP2, expressed in E. coli (Q969E1)

Amino Acid Sequence:
MRNKKFELGL EFPNLPYYID GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL DIRYGVSRIA YSKDFETLKV DFLSKLPEML KMFEDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK KRIEAIPQID KYLKSSKYIA WPLQGWQATF GGGDHPPKSD GSTSGSGHHH HHHSAGLVPR GSTAIGMKET AAAKFERQHM DSPDLGTLEV LFQGPLGSEF-SPIPEVSS AKRRPRRMTP FWRGVSLRPI GASCRDDSEC ITRLCRKRRC SLSVAQ

Predicted Molecular Weight:
33.2kD

Predicted Isoelectric Point:
8.8

Subcellular Location:
Secreted

Accurate Molecular Mass:
39kD as determined by SDS-PAGE reducing conditions
Note: The possible reasons that the actual band size differs from the predicted are as follows:
1. Splice variants: Alternative splicing may create different sized proteins from the same gene
2. Relative charge: The composition of amino acids may affect the charge of the protein.
3. Post-translational modification: Phosphorylation, glycosylation, methylation etc.
4. Post-translation cleavage: Many proteins are synthesized as pro-proteins and then cleaved to give the active form.
5. Polymerization of the target protein: Dimerization, multimerization etc.

Applications:
Suitable for use in ELISA, Western Blot, Immunoprecipitation and SDS-PAGE.
Other applications not tested.

Recommended Dilutions:
Optimal dilutions to be determined by the researcher.

Included Protein Molecular Weight Marker:
168631: Unstained Protein Molecular Weight Marker, 10-70kD
Suppled as a liquid in 62.5mM Tris-H3PO4, pH 7.5, 1mM EDTA, 2% SDS, 100mM DTT, 1mM sodium azide, 0.01% bromo-phenol blue, 33% glycerol. Ready to use; no need to heat, dilute or add reducing agents before use.

Protein Bands: 10kD, 14kD, 18kD, 22kD, 26kD, 33kD, 44kD and 70kD.
Double Intesity Bands: The 26kD, 18kD and 10kD bands are at double intensity to make location and size approximation of proteins easier.

Usage: Allow the marker to reach room temperature and mix thoroughly before use to ensure that any solids that may have precipitated at -20°C will go back into solution. Do not boil!  Load the following volumes on SDS-PAGE gel:
Mini gels: 3-5ul/well
Large gel: 7ul/well
The loading volumes listed above are recommended for gels with thickness of 0.75-1mm. The loading volume should be doubled for 1.5mm thick gels.
Store at -20°C Stable for 6 months after receipt.

Storage and Stability:
Lyophilized powder may be stored at -70°C. Stable for 6 months after receipt. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -70°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Note:
Thermal stability is described by the loss rate of the target protein. The loss rate was determined by accelerated thermal degradation by incubating the protein at 37°C for 48h, with no obvious degradation or precipitation observed. Protein loss is <5% within the expiration date under appropriate storage conditions.