IL-10RB Recombinant Rabbit Monoclonal Antibody [PSH19-60] - BSA and Azide free

Western blot analysis of IL-10RB on different lysates with Rabbit anti-IL-10RB antibody (HA751723) at 1/5,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 cell lysate (no heat)
Lane 3: HDLM-2 cell lysate
Lane 4: HUVEC cell lysate

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 37 kDa
Observed band size: 55-75 kDa

Exposure time: 40 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751723) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Western blot analysis of IL-10RB on different lysates with Rabbit anti-IL-10RB antibody (HA751723) at 1/5,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 cell lysate treated with deglycosylation

Lysates/proteins at 20 µg/Lane.

Predicted band size: 37 kDa
Observed band size: 35-75 kDa

Exposure time: 25 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751723) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of THP-1 cells labeling IL-10RB with Rabbit anti-IL-10RB antibody (HA751723) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IL-10RB antibody (HA751723) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of THP-1 cells labeling IL-10RB.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751723, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).