Antibody Type: Rabbit Polyclonal

Application: FCM

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

IgG

Human,Mouse,Rat (predicted:Cow)

1. The figure annotation:
The blue histogram is unstained cells.
The Orange histogram is cells stained with Rabbit IgG/FITC
The green histogram is cells stained with Rabbit Anti-HMGB1/FITC Conjugated antibody (TMAB-00870-FITC).
Controls
Positive control: HepG 2 cells
Isotype control: Cell lines treated with Rabbit IgG/FITC. instead of the primary antibody to confirm that primary antibody binding is specific. 5 μgin 100 μL 1 X PBS containing 0.5% BSA.
2. Positive control: HepG2 cells
Concebtration: 5 μg/10^6 cells
Incubation conditions: Avoid light , 30 minutes on the ice.
3. Rat splenocytes stained with Anti-HMGB1 Polyclonal Antibody, A488 Conjugated (TMAB-00870-A488) at 1:50.
4. Blank control (blue line): MCF7 (blue).
Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (TMAB-00870)
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.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (TMAB-00870)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The 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 were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

FCM

FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human HMGB1

Human

3146

HMGs,HMG Box,SARS Coronavirus,hmgb1