Anti-HLA-DOB antibody [DOB.L1]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DOB antibody [DOB.L1] (AB312858)

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling HLA-DOB with ab312858 at 1/20000 (0.053 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human skeletal muscle. The section was incubated with ab312858 for 30 mins at room temperature, followed by anti-mouse IgG2b antibody (ab190482) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DOB antibody [DOB.L1] (AB312858)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling HLA-DOB with ab312858 at 1/20000 (0.053 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab312858 for 30 mins at room temperature, followed by anti-mouse IgG2b antibody (ab190482) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-HLA-DOB antibody [DOB.L1] (AB312858)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The 30 kDa band is expected to be the glycoslated form, and the 28 kDa band the deglycosylated form. The identities of the higher MW bands between 50 and 150 kDa are unknown. Exposure time : 3 minutes

All lanes:

Western blot - Anti-HLA-DOB antibody [DOB.L1] (ab312858) at 1/1000 dilution

All lanes:

Human tonsil tissue lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 28 kDa,30 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-HLA-DOB antibody [DOB.L1] (AB312858)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : MOLT-4 (PMID : 2998758). HLA-DOB detected a shift from 30kDa to 28kDa band after deglycosylation treatment. The identities of the higher MW bands between 50 and 150 kDa are unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 125 seconds

All lanes:

Western blot - Anti-HLA-DOB antibody [DOB.L1] (ab312858) at 1/1000 dilution

Lane 1:

Untreated Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate treated with Peptide:N-glycosidase F (PNGase F) at 20 µg

Lane 3:

Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 30 kDa

false

Exposure time: 125s