Western blot analysis of extracts from various cell lines using PHB2 (E1Z5A) Rabbit mAb.
Immunoprecipitation of PHB2 from HeLa cell extracts using using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or PHB2 (E1Z5A) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PHB2 (E1Z5A) Rabbit mAb.
Confocal immunofluorescent analysis of HT-1080 cells using PHB2 (E1Z5A) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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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.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
NOTE: Prepare solutions with Milli-Q or equivalently purified water.
Optional: It may be necessary to perform a lysate pre-clearing step to reduce non-specific binding to the Protein A magnetic beads (See section below).
NOTE: For proteins with molecular weights in the range of around 50 kDa, we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 as a secondary antibody to minimize interference produced by denatured heavy chains. For proteins with molecular weights in the range of around 25 kDa, Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 is recommended to minimize interference produced by denatured light chains.
posted December 2011
protocol id: 125
Protocol Id: 125
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multiwell plates, chamber slides or on coverslips.
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 December 2010
Protocol Id: 32
PHB2 (E1Z5A) Rabbit mAb recognizes endogenous levels of total PHB2 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human PHB2 protein.
The prohibitins PHB1 and PHB2 are highly conserved, multifunctional proteins present in eukaryotic nuclear and mitochondrial compartments (1). Prohibitin-2 (PHB2, REA) was originally identified as an estrogen receptor-specific coregulator. PHB2 directly interacts with hormone-bound estrogen receptor and represses its transcriptional activity through competitive inhibition of Src-1 coactivation of the estrogen receptor (2,3). Together with COUP transcription factors, PHB2 interacts with histone deacetylases HDAC1 and HDAC5 to mediate transcriptional regulation by the estrogen receptor through coupling the deacetylase to the transcription activation complex (4). Prohibitin PHB1/PHB2 heterodimers form large ring complexes on the mitochondrial membrane (5) and act as chaperones to stabilize mitochondrial proteins, such as OPA1 and Hax1, to support mitochondrial morphogenesis and protect against apoptosis (6-8).
Explore pathways + proteins related to this product.
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