Anti-CD90 / Thy1 抗体 [EPR3133]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3133] (AB133350)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD90 / Thy1 with Purified ab133350 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] (AB133350)

Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD90 / Thy1 with Purified ab133350 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] (AB133350)

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] (ab133350) 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

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• WB

Lab

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] (AB133350)

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] (ab133350) 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='/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='/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

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• WB

CiteAb

Western blot - Anti-CD90 / Thy1 antibody [EPR3133] (AB133350)

CD90 / Thy1 western blot using anti-CD90 / Thy1 antibody [EPR3133] ab133350. Publication image and figure legend from Fu, B., Meng, W., et al., 2016, Sci Rep, PubMed 27585821.

ab133350 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab133350 please see the product overview.

GRAMD1A promotes the self-renewal of HCC stem cell and the resistance to chemotherapy.(a) GSEA analysis of the correlation between GRAMD1A levels and tumor recurrence and HCC stem cells gene signature, data set was downloaded from TCGA data set. (b) Hepatosphere formation assay for the effect of GRAMD1A overexpression on self-renewal of HCC stem cells in Huh-7 and HepG2 cells. Representative micrographs (left); quantification of hepatosphere number (right). (c) SP analysis demonstrated the role of GRAMD1A in self-renewal of HCC stem cells by GRAMD1A overexpression. (d) Western blot assay of the expression of HCC stem cells markers CD130 and CD90 by overexpressing or downregulating GRAMD1A in HepG2. (e) Cell viability assay demonstrated the role of GRAMD1A in resistance to Doxorubicin by GRAMD1A overexpression. (f) TUNEL assay for the effect of GRAMD1A overexpression on resistance to Doxorubicin. Representative micrographs (left); quantification of TUNEL positive cell number (right). DAPI was used to stain nucleus. (g) Western blot assay for caspase 3 activity, PARP cleavage and the BCL-XL levels after GRAMD1A overexpression in Huh-7 and HepG2. Data are expressed as mean ± SEM. *p < 0.05.

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