Anti-CD90 / Thy1 antibody [EPR3133] - Low endotoxin, Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3133] - Low endotoxin, Azide free (AB216449)

This data was developed using ab133350, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD90 / Thy1 with Purified ab216449 at 1 : 4000 dilution (0.03 μg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD90 / Thy1 antibody [EPR3133] - Low endotoxin, Azide free (AB216449)

This data was developed using ab133350, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD90 / Thy1 with Purified ab216449 at 1 : 50 dilution (2.44 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Unknown

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] - Low endotoxin, Azide free (AB216449)

This data was developed using ab133350, the same antibody clone in a different buffer formulation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 24116172 and PMID : 30177788).

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] (<a href='/en-us/products/primary-antibodies/cd90-thy1-antibody-epr3133-ab133350'>ab133350</a>) at 1/1000 dilution

Lane 1:

Human glioma lysate at 15 µg

Lane 2:

Human brain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 17 kDa

Observed band size: 25-35 kDa

false

• WB

Lab

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] - Low endotoxin, Azide free (AB216449)

This data was developed using ab133350, the same antibody clone in a different buffer formulation. Anti-THY1 antibody [EPR3133] (ab133350) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133350 was shown to bind specifically to THY1. A band was observed at 25-37 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in THY1 knockout cell line ab262490 (knockout cell lysate ab263925). To generate this image, wild-type and THY1 knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] (<a href='/en-us/products/primary-antibodies/cd90-thy1-antibody-epr3133-ab133350'>ab133350</a>) at 1/2000 dilution

Lanes 1 - 5:

Western blot at 20 µg

Lane 2:

Western blot - Human THY1 (CD90) knockout U-2 OS cell lysate (<a href='/en-us/products/cell-lysates/human-thy1-cd90-knockout-u-2-os-cell-lysate-ab263925'>ab263925</a>)

Lane 2:

Western blot - Human THY1 (CD90) knockout U-2 OS cell line (<a href='/en-us/products/cell-lines/human-thy1-cd90-knockout-u-2-os-cell-line-ab262490'>ab262490</a>)

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 17 kDa

false