Antibody Type: Recombinant Monoclonal

Application: ELISA, WB, IHC, FC, IP

Reactivity: Human

Rabbit IgG

5Y66

Human

1. Western Blot
-Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, THP-1 whole cell lysate, 293 whole cell lysate, A549 whole cell lysate, K562 whole cell lysate, Colo320 whole cell lysate
-All lanes: BAK1 antibody at 0.9μg/ml
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 24, 17 KDa
-Observed band size: 24 KDa
2. IHC image of TMAH-00103 diluted at 1:90 and staining in paraffin-embedded human lymph node tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
3. IHC image of TMAH-00103 diluted at 1:90 and staining in paraffin-embedded human endometrial cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
4. Immunoprecipitating BAK1 in HEK293 whole cell lysate
-Lane 1: Rabbit control IgG instead of TMAH-00103 in HEK293 whole cell lysate.
For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
-Lane 2: TMAH-00103 (3μg) + HEK293 whole cell lysate (500μg)
-Lane 3: HEK293 whole cell lysate (20μg)
5. Overlay histogram showing Hela cells stained with TMAH-00103 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1/200 dilution for 1 h at 4°C. Control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.

ELISA, WB, IHC, FC, IP

WB:1:500-1:5000; IHC:1:50-1:200; IP:1:200-1:1000.

Monoclonal

Unconjugated

A synthetic peptide: Human BAK1

Human

578

Cell Biology