Antibody Type: Recombinant Rabbit Monoclonal

Application: ELISA, WB, IHC, IF, FCM

Reactivity: Human

Rabbit IgG

7Q417

Human

1. Western Blot
-Positive WB detected in: Hela whole cell lysate, U251 whole cell lysate, PC3 whole cell lysate, MCF-7 whole cell lysate
-All lanes: PKN2 antibody at 1:2000
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 113, 117, 112 kDa
-Observed band size: 130 kDa
2. IHC image of TMAH-00986 diluted at 1:100 and staining in paraffin-embedded human colon 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 Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.
3. Immunofluorescence staining of MCF-7 cell with TMAH-00986 at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 509-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
4. Overlay Peak curve showing MCF7 cells stained with TMAH-00986 (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10^6 cells) for 45min at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG(H+L) at 1:200 dilution for 35min at 4°C.Control antibody (green line) was rabbit IgG (1ug/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.

ELISA, WB, IHC, IF, FCM

WB:1:500-1:2000; IHC:1:50-1:200; IF:1:50-1:200; FCM:1:50-1:200.

Monoclonal

Unconjugated

A synthetic peptide: Human PKN2

Human

5586

Signal transduction