[IF=9.207] Li Yin. et al. G6PD activation in TNBC cells induces macrophage recruitment and M2 polarization to promote tumor progression. CELL MOL LIFE SCI. 2023 Jun;80(6):1-20  WB ;  Human.  

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[IF=8.897] Liu, Houli. et al. Immunomodulatory hybrid bio-nanovesicle for self-promoted photodynamic therapy. Nano Res. 2022 Feb;:1-10  WB ;  Mouse.  

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[IF=3.457] Zheng X et al. Dendritic cells and Th17/Treg ratio play critical roles in pathogenic process of chronic obstructive pulmonary disease. (2018) Biomedicine & Pharmacotherapy.108,1141–1151.  IHC ;  Human.  

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Sample:
HL-60(Human) Cell Lysate at 40 ug
Primary: Anti-CD80 (bs-1479R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 32 kD
Observed band size: 57 kD

Sample:
Lane 1: Raw264.7 (Mouse) Cell Lysate at 30 ug
Lane 2: SP2/0 (Mouse) Cell Lysate at 30 ug
Lane 3: Thymus (Rat) Lysate at 40 ug
Lane 4: Raji (Human) Cell Lysate at 30 ug
Lane 5: K562 (Human) Cell Lysate at 30 ug
Primary: Anti-CD80 (bs-1479R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60-65 kD
Observed band size: 60 kD

Blank control: U937(blue).
Primary Antibody: Rabbit Anti-CD80 antibody(bs-1479R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG (orange) ,used under the same conditions.
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min).Primary antibody (bs-1479R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

Blank control:Raji.
Primary Antibody (green line): Rabbit Anti-CD80 antibody (bs-1479R)
Dilution: 2ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Protocol
The cells were 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. Acquisition of 20,000 events was performed.