Antibody Type: Recombinant Rabbit Monoclonal

Application: ELISA, WB, IHC, FCM, IP

Reactivity: Human

Rabbit IgG

9S687

Human

1. Western Blot
-Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, K562 whole cell lysate, HL60 whole cell lysate, PC-3 whole cell lysate
-All lanes: TOP1 antibody at 1:2000
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 91 kDa
-Observed band size: 91 kDa
2. IHC image of TMAH-01197 diluted at 1:100 and staining in paraffin-embedded human small intestine 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 Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
3. IHC image of TMAH-01197 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 IgG polymer labeled by HRP and visualized using 0.05% DAB.
4. Overlay histogram showing HepG2 cells stained with TMAH-01197 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followedby the antibody (1µg/1*10^6 cells) for 1 h at 4°C.The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4°C. Control antibody (green line) was Rabbit IgG (1µg/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.
5. Immunoprecipitating TOP1 in K562 whole cell lysate
-Lane 1: Rabbit control IgG instead of TMAH-01197 in K562 whole cell lysate.
For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
-Lane 2: TMAH-01197(2µg)+ K562 whole cell lysate(500µg)
-Lane 3: K562 whole cell lysate (10µg)

ELISA, WB, IHC, FCM, IP

WB:1:500-1:5000; IHC:1:50-1:200; FCM:1:20-1:200; IP:1:200-1:1000.

Monoclonal

Unconjugated

A synthetic peptide: Human TOP1

Human

7150

Epigenetics and Nuclear Signaling, Cancer