Anti-PHB2 Polyclonal Antibody is a Rabbit antibody targeting PHB2. Anti-PHB2 Polyclonal Antibody can be used in WB,ICC/IF,FCM.

IgG

Human,Mouse,Rat (predicted:Cow,Horse,Sheep)

1. Tissue/cell: colonic epithelial cells;
Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-REA Polyclonal Antibody, Unconjugated (TMAB-10257) 1:100, overnight at 4℃, followed by conjugation to the secondary antibody and DAB staining
2. Protein: spleen (mouse) lysate at 40 μg;
Primary: rabbit Anti-REA (TMAB-10257) at 1:300;
Secondary: HRP conjugated Goat-Anti-rabbit IgG at 1: 5000;
Predicted band size: 33 kD
Observed band size: 35 kD
3. Sample:
Lane 1: Mouse Kidney tissue lysates
Lane 2: Mouse Testis tissue lysates
Lane 3: Mouse NIH/3T3 cell lysates
Lane 4: Rat Kidney tissue lysates
Lane 5: Rat Testis tissue lysates
Lane 6: Human HeLa cell lysates
Lane 7: Human 293T cell lysates
Lane 8: Human HT-1080 cell lysates
Lane 9: Human A431 cell lysates
Lane 10: Human A549 cell lysates
Primary: Anti-PHB2 (TMAB-10257) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 33 kDa
Observed band size: 35 kDa
4. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (PHB2) polyclonal Antibody, Unconjugated (TMAB-10257) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
5. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-PHB2 antibody (TMAB-10257)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

WB: 1:500-2000; ICC/IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated synthetic peptide: human PHB2

Human

PHB2

Prohibitin-2

Acts as a mediator of transcriptional repression by nuclear hormone receptors via recruitment of histone deacetylases. Functions as an estrogen receptor (ER)-selective coregulator that potentiates the inhibitory activities of antiestrogens and represses the activity of estrogens. Competes with NCOA1 for modulation of ER transcriptional activity. Probably involved in regulating mitochondrial respiration activity and in aging.