Anti-CD90 / Thy1 antibody [EPR3132]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labelling CD90 / Thy1 with ab92574 at 1/400 dilution (0.26 μg/ml) followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody at a ready to use concentration. Positive staining on the stroma in human colon carcinoma. The section was incubated with ab92574 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

ab92574 showing positive staining in Normal human tonsil vessels tissue.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

ab92574 showing positive staining in human Lung adenocarcinoma tissue.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

ab92574 showing positive staining in human Skeletal muscle vessels tissue.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

ab92574 showing positive staining in human Cervical carcinoma tissue.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

ab92574 at 1/100 dilution staining CD90 / Thy1 in (1) Human breast carcinoma, (2) Human glioma & (3) Human brain tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue. Detection : DAB staining.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CD90 / Thy1 with ab92574 at 1/400 dilution (0.26 μg/ml) followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody at a ready to use concentration. Positive staining on human tonsil. The section was incubated with ab92574 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

• WB

Lab

Western blot - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

False colour image of Western blot : Anti-CD90 / Thy1 antibody [EPR3132] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab92574 was shown to bind specifically to CD90 / Thy1. A band was observed at 35-45 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD90 / Thy1 antibody [EPR3132] (ab92574) at 1/500 dilution

Lane 1:

Wild-type U-2 OS cell lysate 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>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 17 kDa

Observed band size: 35-45 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

Anti-THY1 antibody [EPR3132] (ab92574) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab92574 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 [EPR3132] (ab92574) at 1/1000 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

• WB

Unknown

Western blot - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

All lanes:

Western blot - Anti-CD90 / Thy1 antibody [EPR3132] (ab92574) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 17 kDa

false

• OI-RD Scanning

Supplier Data

OI-RD Scanning - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.