Fetuin A Recombinant Rabbit Monoclonal Antibody [PSH20-01]

Western blot analysis of Fetuin A on different lysates with Rabbit anti-Fetuin A antibody (HA724121) at 1/20,000 dilution.

Lane 1: Human plasma lysate (40 µg/Lane)
Lane 2: Human serum lysate (40 µg/Lane)

Predicted band size: 39 kDa
Observed band size: 39-55 kDa

Exposure time: 2 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 (HA724121) at 1/20,000 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 HepG2 cells labeling Fetuin A with Rabbit anti-Fetuin A antibody (HA724121) at 1/200 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-Fetuin A antibody (HA724121) at 1/200 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 HepG2 cells labeling Fetuin A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724121, 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).

Fetuin A was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA724121 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724121 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HepG2 cell lysate (input)
Lane 2: HA724121 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of HA724121 in HepG2 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 2 seconds; ECL: K1801

Western blot analysis of Fetuin A on different lysates with Rabbit anti-Fetuin A antibody (HA724121) at 1/100,000 dilution.

Lane 1: Human plasma lysate
Lane 2: Human plasma lysate treated with deglycosylation

Lysates/proteins at 40 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 35-55 kDa

Exposure time: 4 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 (HA724121) at 1/100,000 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.