Confocal immunofluorescent analysis of COLO 205 cells (left, positive) and A-204 cells (right, negative) using HNF1α (E3H9V) Rabbit mAb (IF Specific) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).Learn more about how we get our images.
Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our #12727 Immunofluorescence Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 24
|Immunofluorescence (Immunocytochemistry)||1:400 - 1:1600|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
HNF1α (E3H9V) Rabbit mAb (IF Specific) recognizes endogenous levels of total HNF1α protein. This antibody does not cross-react with HNF1β protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala298 of human HNF1α protein.
Hepatocyte nuclear factor 1α (HNF1α, also known as TCF1 or MODY3) is a transcription factor that plays a role in the tissue-specific regulation of liver gene expression (1). Research has shown that heterogeneous mutations of HNF1α are linked to maturity onset diabetes of the young (MODY) (2). Recent studies indicate that increased concentrations of free fatty acids can reduce the expression of FoxA2/HNF3β and HNF1α in pancreatic β-cells and lead to their nuclear exclusion, resulting in symptoms of several metabolic syndromes (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
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