Antibody Type: Recombinant Rabbit Monoclonal

Application: ELISA, WB, IHC, IF, FCM

Reactivity: Human

Rabbit IgG

4W795

Human

1. Western Blot
-Positive WB detected in: Ntera-2 whole cell lysate, Hela whole cell lysate, A549 whole cell lysate, HepG2 whole cell lysate, PC3 whole cell lysate, U87 whole cell lysate
-All lanes: HIF1AN antibody at 1:2000
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 41 kDa
-Observed band size: 36-53 kDa
2. IHC image of TMAH-00544 diluted at 1:100 and staining in paraffin-embedded human testis 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 polymer IgG labeled by HRP and visualized using 0.05% DAB.
3. IHC image of TMAH-00544 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.
4. Immunofluorescence staining of HepG2 cell with TMAH-00544 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 525-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
5. Overlay Peak curve showing Hela cells stained with TMAH-00544 (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 HIF1AN

Human

55662

Epigenetics and Nuclear Signaling, Cancer, Cardiovascular