Antibody Type: Rabbit Polyclonal

Application: WB,FCM

Reactivity: Human (predicted:Mouse,Rat,Chicken)

IgG

Human (predicted:Mouse,Rat,Chicken)

1. Sample:
Lane 1: Hela (Human) Cell Lysate at 30 μg
Lane 2: A431 (Human) Cell Lysate at 30 μg
Lane 3: U251 (Human) Cell Lysate at 30 μg
Lane 4: HepG2 (Human) Cell Lysate at 30 μg
Primary: Anti-HIF-1 Alpha (TMAB-00857) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 92 kDa
Observed band size: 120 kDa
2. Blank control (blue line): Hela (fixed with 80% methanol (5 min at-20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-HIF-1 Alpha antibody (TMAB-00857),
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
3. Sample:
Lane 1: Recombinant human HIF-1 Alpha protein, N-Trx-His
Primary: Anti-HIF-1 Alpha (TMAB-00857) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 92 kDa
Observed band size: 52 kDa
4. The HepG2 (H) （treated with 500uM CoCl2 for 6 hours) cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C,the cells then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at room temperature). Primary Antibody (green): Rabbit Anti-HIF-1 Alpha antibody (TMAB-00857): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC: 1 μg/test. Isotype Control (orange): Rabbit IgG. Blank control (black): PBS.

WB,FCM

WB: 1:500-2000; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human HIF-1 Alpha

Human

3091

HIF,Hydroxylases,HIF,Jumonji containing proteins,Methylation