Flt-3/Flk-2/CD135 Overexpression Lysate Summary
Description |
Flt-3/Flk-2/CD135 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC211459
Accession#: NM_004119
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
FLT3 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Theoretical MW |
112.7 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. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for Flt-3/Flk-2/CD135 Overexpression Lysate
Background
Stem cell tyrosine kinase (STK-1) has been cloned from a CD34+ hematopoietic stem cell enriched library and identified as the human homolog of a previously identified gene of mouse origin designated either Flk-2 or Flt-3. The STK-1 cDNA encodes a protein of 993 amino acids with 85% identity to Flt-3/Flk-2. STK-1 is a member of the type III receptor tyrosine kinase family that includes Kit (steel factor receptor), Fms and PDGF. STK-1 expression in blood and marrow is restricted to CD34+ cells, a population greatly enriched for hematopoietic stem/progenitor cells. STK-1 antiserum recognizes two polypeptides in these cells. The mouse homolog of STK-1, designated Flt-3/ Flk-2, is expressed at high levels in hematopoietic cells and also in neural, gonadal, hepatic and placental tissues. It has been suggested that STK-1 and its murine homolog Flt-3/ Flk-2 may function as growth factor receptors on hematopoietic stem and/or progenitor cells.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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