mouse anti-wt1 antibody clone 6f-h2  (Cell Marque)

Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: WT1 protein expression in tissues at different stages of multiple myeloma (MM): Rates of WT1 protein expression in MM tissues at different clinical settings (A) , Rates of cytoplasmic and nuclear WT1 staining in different tissue samples (B) , Histology score (H-score) of WT1 cytoplasmic staining (C) , and nuclear staining (D) in different myeloma samples at first diagnosis or relapse stage (* p < 0.05, ** p < 0.01).

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Expressing, Staining, Biomarker Discovery

Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: Immunohistochemistry (IHC) staining of WT1 protein: (A–D) show a representative cytoplasmic staining with different intensities in samples: Negative staining (A) , + (B) , ++ (C) and +++ (D) ; (E–H) show a representative nuclear staining with different intensities in samples: Negative staining (E) , + (F) , ++ (G) and +++ (H) .

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Immunohistochemistry, Staining, Negative Staining

Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: Characteristics of WT1 positivity in samples.

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Staining, Isolation

Journal: International Journal of Molecular Sciences

Article Title: The Detection of Immunity against WT1 and SMAD4 P130L of EpCAM + Cancer Cells in Malignant Pleural Effusion

doi: 10.3390/ijms232012177

Figure Lengend Snippet: WT1 expression on EpCAM + cancer cells and detection of memory subsets of WT1-CTLs in MPE samples. ( A ) Expression of WT1 on EpCAM positive or negative cells sorted from whole cells in MPE 1st . PC, phase contrast; WT1 cells were stained with anti-WT1 antibody, followed by Alexa Fluor® Plus 488 secondary antibody; DNA, DAPI staining. The white bar indicates 100 μm. ( B ) The ratio of WT-CTLs is defined as WT1 tetramer + CD8 + T cells to MPE samples 1st or MPE 2nd (upper panel). The memory subsets of WT1-CTLs during disease progression (middle panel). CD62L + CD45RO + : central memory T cells (T CM ); CD62L + CD45RO − : naïve T cells (T N ); CD62L − CD45RO + : effector memory T cells (T EM ), CD62L − CD45RO − : terminal effector T cells (T TE ). The function of WT1-CTLs in MPE was evaluated by ELISpot assay for IFN-γ secretion under WT1 235 peptide stimulation (lower panel). The mean number of spots of WT1 235 peptide-specific was shown. The error bar indicated the mean and standard deviation. Unpaired t -test, * p < 0.05.

Article Snippet: Subsequently, 1 × 10 5 cells were fixed with a 10% formalin neutral buffer solution (Wako Pure Chemicals Ltd., Osaka, Japan) for 30 min and then permeabilized with a 0.1% Triton X-100 (Sigma-Aldrich Co. LLC, St. Louis, MO, USA) solution in PBS at room temperature for 5 min and blocked with UltraCruz Blocking Reagent (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) for 60 min. After blocking, the cells were incubated with a mouse monoclonal primary anti-WT1 antibody (1:100, clone 6F-H2, Thermo Fisher Scientific, Waltham, MA, USA) at 4 °C for 16 h. Subsequently, a goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody conjugated with Alexa Fluor plus 488 (1:200, Thermo Fisher Scientific) was added at room temperature for 30 min.

Techniques: Expressing, Staining, Biomarker Discovery, Enzyme-linked Immunospot, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: The Detection of Immunity against WT1 and SMAD4 P130L of EpCAM + Cancer Cells in Malignant Pleural Effusion

doi: 10.3390/ijms232012177

Figure Lengend Snippet: Detection of CD8 + T cell response to HLA-A*11:01 restricted SMAD4 P130L neoantigen in MPE. ( A ) WT1 tetramer-CD8 + T cells sorted from MPE 1st were expanded with HPL-IFN-DCs containing each predicted neoantigen peptide for SMAD4 P130L . For ELISpot assay to detect IFN-γ production after in vitro expansion, these cells were stimulated using each SMAD4 P130L peptide. Reactivity was shown in the box plot (per each well in 96 plates). DMSO was used as a negative control. A Wilcoxon signed-rank test was performed, * p < 0.05. ( B ) CD8 + T cells purified from in vitro expanded cells with HPL-IFN-DCs contained SMAD4-Neo1 peptides in A were stimulated by HEV0271 cells (HLA-A*11:01 homozygous) containing SMAD4-Neo1 (SVCVNLYH) or SMAD4-WT1 (SVCVNPYH). The reactivity to IFN-γ was evaluated by ELISpot assay. The error bar indicated the mean and standard deviation. Dunnett’s test, * p < 0.05.

Article Snippet: Subsequently, 1 × 10 5 cells were fixed with a 10% formalin neutral buffer solution (Wako Pure Chemicals Ltd., Osaka, Japan) for 30 min and then permeabilized with a 0.1% Triton X-100 (Sigma-Aldrich Co. LLC, St. Louis, MO, USA) solution in PBS at room temperature for 5 min and blocked with UltraCruz Blocking Reagent (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) for 60 min. After blocking, the cells were incubated with a mouse monoclonal primary anti-WT1 antibody (1:100, clone 6F-H2, Thermo Fisher Scientific, Waltham, MA, USA) at 4 °C for 16 h. Subsequently, a goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody conjugated with Alexa Fluor plus 488 (1:200, Thermo Fisher Scientific) was added at room temperature for 30 min.

Techniques: Enzyme-linked Immunospot, In Vitro, Negative Control, Purification, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: The Detection of Immunity against WT1 and SMAD4 P130L of EpCAM + Cancer Cells in Malignant Pleural Effusion

doi: 10.3390/ijms232012177

Figure Lengend Snippet: Memory subsets of WT1 tetramer − CD8 + T cells and detection of SMAD4 P130L -specific CD8 + T cell expanded from MPE samples. ( A ) WT1 tetramer − CD8 + T cells in MPE 1st or MPE 2nd were stained with antibodies against CD62L and CD45RO for memory T cell subsets. ( B ) WT1 tetramer − CD8 + T cells were sorted from MPE 1st or MPE 2nd , expanding in vitro with HPL-IFN-DCs containing SMAD4-Neo1 peptide. These cells were stimulated with SMAD4-Neo1 or DMSO to detect IFN-γ-producing cells by ELISpot assay. The ratio of IFN-γ spots of SMAD4-Neo1 to control was shown in the box plot (per each well in 96 plates). Mann–Whitney U test, * p < 0.05.

Article Snippet: Subsequently, 1 × 10 5 cells were fixed with a 10% formalin neutral buffer solution (Wako Pure Chemicals Ltd., Osaka, Japan) for 30 min and then permeabilized with a 0.1% Triton X-100 (Sigma-Aldrich Co. LLC, St. Louis, MO, USA) solution in PBS at room temperature for 5 min and blocked with UltraCruz Blocking Reagent (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) for 60 min. After blocking, the cells were incubated with a mouse monoclonal primary anti-WT1 antibody (1:100, clone 6F-H2, Thermo Fisher Scientific, Waltham, MA, USA) at 4 °C for 16 h. Subsequently, a goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody conjugated with Alexa Fluor plus 488 (1:200, Thermo Fisher Scientific) was added at room temperature for 30 min.

Techniques: Staining, In Vitro, Enzyme-linked Immunospot, Control, MANN-WHITNEY

Journal: Disease Models & Mechanisms

Article Title: Circulating fibroblast growth factor-2 precipitates HIV nephropathy in mice

doi: 10.1242/dmm.048980

Figure Lengend Snippet: rAd-FGF-2 induced de-differentiation changes in podocytes and albuminuria and increased the blood urea nitrogen (BUN) levels of HIV-Tg 26 mice. (A-H) Representative immunohistochemistry staining for WT1 and synaptopodin (both brown in renal sections harvested 14 days after the adenoviral injections) ( n =4-5 mice per group). Scale bar: 20 µm. (I,J) Percentage changes in WT1 + (I) and synaptopodin + (J) cells relative to the corresponding control groups (mean±s.e.m.; n =4-5 mice per group). (K) WT and HIV-Tg 26 mice injected with rAd-FGF-2 vectors developed significant albuminuria compared to the corresponding control groups ( n =4-5 mice per group). (L) HIV-Tg 26 mice showed elevated BUN levels 28 days after the injection of rAd-FGF-2 vector ( n =4-5 mice per group). Statistical significance was determined using a Mann–Whitney unpaired t- test. * P <0.05, ** P <0.01 and *** P <0.001, compared to the corresponding LacZ groups.

Article Snippet: WT-1 staining was assessed with a mouse monoclonal anti-human WT1 antibody (clone 6F-H2, Millipore Sigma; 1:600 dilution).

Techniques: Immunohistochemistry, Staining, Control, Injection, Plasmid Preparation, MANN-WHITNEY