Anti-HMGB1 Polyclonal Antibody is a Rabbit antibody targeting HMGB1. Anti-HMGB1 Polyclonal Antibody can be used in FCM.

IgG

Human,Rat (predicted:Mouse,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: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human HMGB1

Human

HMGB1

High mobility group protein B1

HMGs,HMG Box,SARS Coronavirus,hmgb1

DNA binding proteins that associates with chromatin and has the ability to bend DNA. Binds preferentially single-stranded DNA. Involved in V(D)J recombination by acting as a cofactor of the RAG complex. Acts by stimulating cleavage and RAG protein binding at the 23 bp spacer of conserved recombination signal sequences (RSS). Heparin-binding protein that has a role in the extension of neurite-type cytoplasmic processes in developing cells.