resource source identifier antibodies α stn b72 3 santa cruz biotechnology cat  (Santa Cruz Biotechnology)

Journal: Virulence

Article Title: Eukaryotic translation elongation factor 1 alpha 1 facilitates coxsackievirus B replication through interacting with PABP, viral protein 3 CD and viral RNA

doi: 10.1080/21505594.2025.2563925

Figure Lengend Snippet: CVB replication requires eEF1A1 participation. (A-F) HeLa cells were transfected with either pEGFP-eEF1A1 or pEGFP-eEF1A2. Control cells were transfected with empty vector. Cells were infected by CVB3 (MOI = 1) at 24 h of post-transfection. Cells were collected at 12 h of post-infection and analyzed by RT-qPCR (C and F) and immunoblotting (A and D). (G-L) HeLa cells were transfected with either si-eEF1A1 or si-eEF1A2 for 24 h. Control cells were transfected with scramble RNA (si-control). Cells were then infected with CVB3 (MOI = 1) for 12 h and analyzed by RT-qPCR (I and L) and immunoblotting (G and J). Quantifications of Western blots (B, E, H, K) were performed with ImageJ. Ctrl: control; * p < 0.05; ** p < 0.01; **** p < 0.0001; ns: non-significant. Data represent means ± s.D. Of n = 3 biologically independent experiments.

Article Snippet: The antibodies used in this study were listed below: eEF1A1 polyclonal antibody (11402–1-AP, Proteintech, Wuhan, China), eEF1A2 polyclonal antibody (16091–1-AP, Proteintech, Wuhan, China), GAPDH monoclonal antibody (60004–1-AP, Proteintech, Wuhan, China), EGFP tag polyclonal antibody (50430–2-AP, Proteintech, Wuhan, China), DYKDDDDK tag polyclonal antibody (Binds to Flag tag epitope; 20543–1-AP, Proteintech, Wuhan, China), NF-κB p65 polyclonal antibody (10745–1-AP, Proteintech, Wuhan, China), Phospho-NF-κB p65 (Ser536) antibody (3031, Cell Signaling, Boston, MA), MYC tag polyclonal antibody (16286–1-AP, Proteintech, Wuhan, China), PABP polyclonal antibody (10970–1-AP, Proteintech, Wuhan, China), PCBP2 polyclonal antibody (15070–1-AP, Proteintech, Wuhan, China).

Techniques: Transfection, Control, Plasmid Preparation, Infection, Quantitative RT-PCR, Western Blot

Journal: Virulence

Article Title: Eukaryotic translation elongation factor 1 alpha 1 facilitates coxsackievirus B replication through interacting with PABP, viral protein 3 CD and viral RNA

doi: 10.1080/21505594.2025.2563925

Figure Lengend Snippet: eEF1A1 expression is upregulated by NF-kB in CVB3-infected cells. (A-C) HeLa cells were infected with CVB3 (MOI = 1). Cells were collected at 0, 3, 6, 9, and 24 h of post-infection. The RNA levels of CVB, eEF1A1, and eEF1A2 were detected by RT-qPCR. (D and E) HeLa cells were infected with CVB3 (MOI = 1) for 12 h. Cells were collected and subjected to immunoblotting to determine the expression levels of phospho-NF-κB p65, NF-κB p65, and 3D of CVB3. (F-I) HEK293T cells were transfected with pmCherry-p65 or empty vector (pmCherry-HA) for 24 h. Cells then were infected with CVB3 (MOI = 1) for 12 h. The expression of eEF1A1 was determined by RT-qPCR and immunoblotting. Quantification of the blots was performed with ImageJ. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns: non-significant. Data represent means ± s.d. of n = 3 biologically independent experiments.

Article Snippet: The antibodies used in this study were listed below: eEF1A1 polyclonal antibody (11402–1-AP, Proteintech, Wuhan, China), eEF1A2 polyclonal antibody (16091–1-AP, Proteintech, Wuhan, China), GAPDH monoclonal antibody (60004–1-AP, Proteintech, Wuhan, China), EGFP tag polyclonal antibody (50430–2-AP, Proteintech, Wuhan, China), DYKDDDDK tag polyclonal antibody (Binds to Flag tag epitope; 20543–1-AP, Proteintech, Wuhan, China), NF-κB p65 polyclonal antibody (10745–1-AP, Proteintech, Wuhan, China), Phospho-NF-κB p65 (Ser536) antibody (3031, Cell Signaling, Boston, MA), MYC tag polyclonal antibody (16286–1-AP, Proteintech, Wuhan, China), PABP polyclonal antibody (10970–1-AP, Proteintech, Wuhan, China), PCBP2 polyclonal antibody (15070–1-AP, Proteintech, Wuhan, China).

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Transfection, Plasmid Preparation

Journal: Cell Reports Medicine

Article Title: Pan-carcinoma sialyl-Tn-targeting expands CAR therapy to solid tumors

doi: 10.1016/j.xcrm.2025.102350

Figure Lengend Snippet: AM52.1 antibody shows specific binding to sialyl-Tn (A) Schematic illustration of the predominant glycan structures in ovine submaxillary mucin (OSM) and its desialylated form, Asialo-OSM (A-OSM). (B) Binding of AM mAbs to varying concentrations of OSM and A-OSM, measured by indirect enzyme-linked immunosorbent assay (ELISA). B72.3, CC49, and 1E3 antibodies were included as controls. (C) Binding specificity of AM mAbs to various sialyl-Tn (STn)-related O -glycan structures, as measured by glycan microarray analysis; 10 μg/mL of each antibody were used. B72.3 mAb served as a control for the assay. Refer to A for a comprehensive list of the analyzed glycan structures. (D) Gastrointestinal cancer wild-type (WT) and core 1 β1,3-galactosyltransferase (C1GalT1) knockout (KO) cells were stained for flow cytometry using the AM mAbs, and well-established anti-STn and anti-Tn antibodies as controls. Histograms represent the cell populations stained with the annotated antibodies. (E) Cell lines of different origins (lymphoma, osteosarcoma, melanoma, glioblastoma, prostate, colorectal, gastric, ovarian, breast, and oral cancer) were stained with AM52.1. Representative flow cytometry plots of isotype control and AM52.1 staining are shown. (F) AM51.1 and AM52.1 antibodies, alongside with the positive control B72.3, were used to stain tissue from gastric, colon, pancreatic, prostate, and ovarian carcinomas. Sections show representative staining of tumor tissue with the annotated antibodies. Scale bar corresponds to 200 μm. (G) Graphs depicting the percentage of carcinoma sections exhibiting positive staining for each antibody tested and the corresponding H-score for each tissue type. (H) Healthy tissues (heart, kidney, liver, spleen, lung, thyroid, lip, vocal cords, cervix, and endometrium) were stained with the AM antibodies and the B72.3 positive control. Scale bar corresponds to 100 μm. (I) Pie chart representing cell population distribution in human peripheral blood mononuclear cells (PBMCs) from healthy donors ( n = 5) stained with the AM mAbs. (J) Histograms depicting the percentage distribution of various immune cell populations stained with AM51.1 and AM52.1. Data are represented as mean ± standard deviation (SD).

Article Snippet: α-STn (CC49) , Santa Cruz Biotechnology , Cat#sc-20043; RRID: N/A.

Techniques: Binding Assay, Glycoproteomics, Indirect ELISA, Enzyme-linked Immunosorbent Assay, Microarray, Control, Knock-Out, Staining, Flow Cytometry, Positive Control, Standard Deviation

Journal: Cell Reports Medicine

Article Title: Pan-carcinoma sialyl-Tn-targeting expands CAR therapy to solid tumors

doi: 10.1016/j.xcrm.2025.102350

Figure Lengend Snippet: AM52.1CAR T cells are effective and specific in a preclinical model of patient-derived organoids (A) sialyl-Tn (STn) expression evaluation using CC49 and AM52.1 mAbs in five patient-derived organoids (PDOs). Scale bars correspond to 100 and 50 μm, as specified. (B) Schematic outlining the experimental design followed to establish the co-culture of the PDOs with the immune cells. (C) Viability of a STn-negative (PDO2) and a STn-positive (PDO5) PDOs upon co-culture with specified CAR T cells at different effector:target (E:T) ratios. Statistical comparison reflects differences at the same E:T ratio between the different organoids. n = 1 healthy donor. (D) Immunofluorescent analysis of cleaved caspase-3 (CC3) expression in the selected PDOs after co-culture with indicated CAR T cells at an E:T ratio of 10:1. Scale bars correspond to 50 and 10 μm, as specified. (E) Viability of PDOs derived from normal gastric mucosa (PDO1) and gastric tumor tissue (PDO2-5) with different levels of STn expression following co-culture with specified CAR T cells. The data correspond to the E:T ratio of 10:1. n = 3 healthy donors. Data are represented as mean ± standard deviation (SD). Statistical significance was determined using paired t test. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001.

Article Snippet: α-STn (CC49) , Santa Cruz Biotechnology , Cat#sc-20043; RRID: N/A.

Techniques: Derivative Assay, Expressing, Co-Culture Assay, Comparison, Standard Deviation

Journal: Functional & Integrative Genomics

Article Title: Integrated Nanopore and short-read RNA sequencing identifies dysregulation of METTL3- m6A modifications in endocrine therapy- sensitive and resistant breast cancer cells

doi: 10.1007/s10142-025-01658-2

Figure Lengend Snippet: Western blot analysis of DEGs with DMR DEGs in LCC9 compared to MCF-7 cells. The R package ggtranscript ( https://dzhang32.github.io/ggtranscript/ ) was used to plot the m6A modifications identified with m6ANet analysis in the indicated transcripts of PDIA6, ACTB, EEF1A2, and FLNA in LCC9 vs. MCF-7 cells A, D, G, J . For the western blots B, E, H, K, six biological replicate samples of WCE from MCF-7 and LCC9 were probed with the indicated antibody and each protein’s abundance was quantified relative to Ponceau S staining of that same blot and then normalized to that protein’s abundance in MCF-7 cells. (C, F, I, L) Results MeRIP RT-qPCR for specific m6A-modified sites in C PDIA6, F ACTB, I EEF1A2, and L FLNA genes in MCF-7 cells treated with 1 μM STM2457 for 24 h. A PDIA6 transcript abundance was higher (log2FC 0.63, q-value < 0.05) and there was more m6A enrichment on the 5 transcripts in LCC9 compared to MCF-7 cells. B PDIA6 protein. D ACTB transcript abundance was downregulated (log2FC −1.18, q-value < 0.05) and there was less m6A enrichment on 5 transcripts in LCC9 compared to MCF-7 cells. E ACTB protein. G EEF1A2 transcript abundance was downregulated (log2FC −2.02, q-value < 0.05) and there was less m6A enrichment on 3 transcripts in LCC9 compared to MCF-7 cells. H EEF1A2 protein. J FLNA transcript abundance was downregulated (log2FC −1.61, q-value < 0.05) and there was more m6A enrichment on 2 transcripts in LCC9 compared to MCF-7 cells. K FLNA protein. DMR = differential modification rate, p-value < 0.001. Individual data are plotted from replicate experiments and error bars represent SEM (n = 5). Western and MeRIP-RT-qPCR data were analyzed using a two-tailed t test in Graph Pad Prism: * p < 0.05, *** p < 0.001, **** p < 0.0001

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies: PDIA6 (#18233–1-AP, ProteinTech, Rosemont, IL, USA), FLNA (#67133–1-Ig, ProteinTech), EEF1A2 (16091–1-AP, ProteinTech), ACTB (#A5316, Sigma Aldrich), and AMIGO2 (#14076–1-AP, ProteinTech).

Techniques: Western Blot, Staining, Quantitative RT-PCR, Modification, Two Tailed Test