mouse monoclonal anti ship1 p1c1  (Santa Cruz Biotechnology)

Journal: STAR Protocols

Article Title: Protocol for gene knockdown using siRNA in primary cultured neonatal murine microglia

doi: 10.1016/j.xpro.2024.102867

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Article Snippet: Mouse monoclonal anti-SHIP1 (P1C1) (1:200 dilution) , Santa Cruz Biotechnology , Cat#sc-8425; RRID: AB_628250.

Techniques: Recombinant, Transfection, Modification, Sterility, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Software, Suspension, Suction Filtration, Membrane, Microscopy

Journal: Cell Reports Medicine

Article Title: Discovery of galectin-8 as an LILRB4 ligand driving M-MDSCs defines a class of antibodies to fight solid tumors

doi: 10.1016/j.xcrm.2023.101374

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Article Snippet: Mouse monoclonal anti-SHIP-1 (Clone P1C1) , Santa Cruz Biotechnology , Cat#sc-8425; RRID: AB_628250.

Techniques: Recombinant, Derivative Assay, Microarray, Enzyme-linked Immunosorbent Assay, Lysis, Blocking Assay, Staining, Transfection, Marker, Flow Cytometry, shRNA, Plasmid Preparation, Negative Control, Silver Staining, Bicinchoninic Acid Protein Assay, Selection, Extraction, Labeling, RNA Expression, Cell Culture, Software

Journal: bioRxiv

Article Title: Disc Large Homolog 1 is critical for early microcluster formation and activation in human T cells

doi: 10.1101/2021.05.24.445526

Figure Lengend Snippet: A and B. Microcluster analysis. Scale bars, 10 μm. Data are representative of three experiments. (A) Jurkat cell analysis. Microclusters of ZAP-70–YFP (green) and phosphorylated TCR-ζ (red) were visualized in immunostained Jurkat cells stably expressing ZAP-70–YFP, pretreated with control or DLG1-specific siRNA. Cells were dropped onto coverslips coated with anti-CD3 (stimulated) or anti-CD45 (unstimulated) and visualized after 5 min. (B) Primary human CD4+ T cells. Microclusters of phosphorylated Y493 ZAP-70 (green) and phosphorylated TCR-ζ (red) were visualized in immunostained human T cells, pretreated with control or DLG1-specific siRNA. Cells were dropped onto coverslips coated with anti-CD3 + anti-CD28 (stimulated) or anti-CD45 (unstimulated) and visualized after 5 min. C and D. Immunoblot analysis of lysates from Jurkat cells (C) and human T cells (D) pretreated with control or DLG1-specific siRNA and then activated with soluble anti-CD3 (monoclonal antibody OKT3) and antibody to mouse immunoglobulin G (IgG), followed by probing for phosphorylated proteins. The membranes were then stripped and reblotted with antibodies to detect total cellular proteins corresponding to the phosphorylated proteins.

Article Snippet: The anti-mouse monoclonal antibodies that were used included, anti TCR-ζ (6B10.2;1239), anti-SAP97 (Dlg1; clone 2D11, sc-9961) and anti-Lck (3A5; 433), siRNAs including control siRNA-A (sc-37007), siRNA plasmid against hDlg1 (sc-36452) and hSHP1 (sc-36490)and agarose-conjugated CD3-ζ mouse monoclonal IgG1 (1239-AC) from Santa Cruz, California,USA.

Techniques: Cell Analysis, Stable Transfection, Expressing, Control, Western Blot

Journal: bioRxiv

Article Title: Disc Large Homolog 1 is critical for early microcluster formation and activation in human T cells

doi: 10.1101/2021.05.24.445526

Figure Lengend Snippet: A. Microclusters of phosphorylated Y493 ZAP-70(green) and phosphorylated TCR-ζ (red) in immunostained wildtype or DLG1 KO Jurkat cells were visualized 5 min after the cells were dropped onto coverslips coated with anti-CD3 (stimulated) or anti-CD45 (unstimulated) and visualized. Scale bars, 10 μm. Data are representative of three experiments. B. Immunoblot analysis of lysates from wildtype and DLG1 KO Jurkat cells activated with soluble anti-CD3 (monoclonal antibody OKT3) and antibody to mouse immunoglobulin G (IgG). Blots were probed for the indicated phosphorylated proteins, then stripped and reprobed for detection of total cellular proteins corresponding to the phosphorylated proteins.

Article Snippet: The anti-mouse monoclonal antibodies that were used included, anti TCR-ζ (6B10.2;1239), anti-SAP97 (Dlg1; clone 2D11, sc-9961) and anti-Lck (3A5; 433), siRNAs including control siRNA-A (sc-37007), siRNA plasmid against hDlg1 (sc-36452) and hSHP1 (sc-36490)and agarose-conjugated CD3-ζ mouse monoclonal IgG1 (1239-AC) from Santa Cruz, California,USA.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Disc Large Homolog 1 is critical for early microcluster formation and activation in human T cells

doi: 10.1101/2021.05.24.445526

Figure Lengend Snippet: A. Microclusters of phosphorylated Y493 ZAP-70 (green) and phosphorylated TCR-ζ (red) were visualized in immunostained wildtype or DLG1 KO Jurkat cells that were pretreated with control or SHP1 specific siRNA. Cells were dropped onto coverslips coated with anti-CD3 (stimulated) or anti-CD45 (unstimulated) and visualized after 5 min. Scale bars, 10 μm. Data are representative of three experiments. B. Immunoblot analysis of lysates from wildtype and DLG1 KO Jurkat cells. Cells were pretreated with control or SHP1-specific siRNA, then activated with soluble anti-CD3 (monoclonal antibody OKT3) and antibody to mouse immunoglobulin G (IgG), followed by probing for phosphorylated proteins. Membrane were then stripped and re-probed for detection of total cellular proteins corresponding to the phosphorylated proteins.

Article Snippet: The anti-mouse monoclonal antibodies that were used included, anti TCR-ζ (6B10.2;1239), anti-SAP97 (Dlg1; clone 2D11, sc-9961) and anti-Lck (3A5; 433), siRNAs including control siRNA-A (sc-37007), siRNA plasmid against hDlg1 (sc-36452) and hSHP1 (sc-36490)and agarose-conjugated CD3-ζ mouse monoclonal IgG1 (1239-AC) from Santa Cruz, California,USA.

Techniques: Control, Western Blot, Membrane

Journal: Nature Communications

Article Title: Loss of NDRG2 expression activates PI3K-AKT signalling via PTEN phosphorylation in ATLL and other cancers

doi: 10.1038/ncomms4393

Figure Lengend Snippet: ( a ) A western blot analysis of NDRG2, PTEN, phosphorylated PTEN (p-PTEN) (Ser370, Ser380/Thr382/Thr383 and Ser385), non-p-PTEN (Ser380/Thr382/Thr383), SHIP1, AKT, p-AKT (Ser473 and Thr308) and Tax was performed in primary leukaemic cells from acute-type ATLL patients. The CD4 + T lymphocytes from healthy volunteers (CD4 + T lymph) served as the controls. No expression of Tax was detected in CD4 + T lymphocytes from healthy volunteers or ATLL cells from acute-type ATLL patients. The graph shows relative band intensity of p-PTEN (Ser380/Thr382/Thr383) and non-p-PTEN (Ser380/Thr382/Thr383). Each p-PTEN or non-p-PTEN value was divided by the total PTEN value for a given sample. The data are expressed as the mean value±s.d. (* P <0.05, Student’s t- test). The data are representative of three experiments. ( b ) A western blot analysis with the same antibodies as in was performed in the T-ALL and ATLL cell lines. Asterisk, nonspecific band. HTLV-1-transformed T-cell lines (HUT102, MT2 and MT4) expressed high levels of Tax. The graph shows relative band intensity of p-PTEN (Ser380/Thr382/Thr383) and non-p-PTEN (Ser380/Thr382/Thr383). Each p-PTEN or non-p-PTEN value was divided by the total PTEN value for a given sample. The data are representative of three experiments. ( c ) Quantitative RT–PCR analysis of PTEN in CD4 + T lymphocytes from five healthy volunteers and ATLL cells from seven acute-type ATLL patients, along with four T-ALL cell lines and eight ATLL cell lines. The relative amounts of mRNA were normalized against β-actin mRNA and expressed relative to the mRNA abundance in healthy control sample 1. The mean±s.d. is shown; * P <0.05; NS, not significant (Mann–Whitney U -test). The data are representative of two experiments.

Article Snippet: Rabbit polyclonal antibodies against phospho-PTEN (Ser370) (07-889) and PI3K p85 (06-195) were obtained from Upstate/Millipore, rabbit polyclonal antibody against phospho-PTEN (Ser385) (44-1064G) was obtained from Biosource, mouse monoclonal antibody against SHIP1 (P1C1) and goat polyclonal antibody against NDRG2 (E20) were obtained from Santa Cruz Biotechnology, rabbit polyclonal antibody against GFP (598) was obtained from MBL, and rat monoclonal antibodies against mouse B220 (RA3-6B2), mouse CD4 (RM4-5) and mouse CD8 (53-6.7) and hamster monoclonal antibody against mouse CD3 (500A2) were obtained from BD Pharmingen.

Techniques: Western Blot, Expressing, Transformation Assay, Quantitative RT-PCR, Control, MANN-WHITNEY