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
PIBF1
Applications/Dilutions
Dilutions
ELISA 2 ug/ml
Western Blot
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
89.6 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.
Reviewed Applications
Read 1 Review rated 4 using NBL1-14389 in the following applications:
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 PIBF1 Overexpression Lysate
C13orf24
chromosome 13 open reading frame 24
KIAA1008
PIBF
PIBF1
progesterone immunomodulatory binding factor 1
progesterone-induced blocking factor 1
progesterone-induced-blocking factor 1
RP11-505F3.1
Background
PIBF (progesterone-induced blocking factor 1) is synthesized during pregnancy in response to progesterone by progesterone receptor-positive T lymphocytes (mostly gamma-delta T cells). In the presence of PIBF, natural killer (NK) cells inhibit the release of perforin from storage granules and, therefore, fail to lyse target cells. In humans, the amount of cells that express PIBF is significantly higher in healthy pregnant women than in women at risk for premature pregnancy termination. PIBF has a predicted 89 kDa molecular mass; the full-length PIBF is associated with the nucleus, whereas secretion of shorter forms, such a 34 kDa protein is induced by activation of the cell. Research suggests that PIBF functions as a transcription factor in its full-length form, while smaller forms may act as cytokines.
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.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.