Journal: PLoS ONE

Article Title: Chondroitin Sulfate Proteoglycan CSPG4 as a Novel Hypoxia-Sensitive Marker in Pancreatic Tumors

doi: 10.1371/journal.pone.0100178

Figure Lengend Snippet: Immunoblotting confirmed the specificity of polyclonal H-300 ( A ) and monoclonal LHM2 antibodies ( B ), generated against recombinant protein and melanoma-derived antigens, respectively. The images demonstrate differences in size between pancreatic (band-2) and melanoma (SK-MEL-28, band-1) antigens, but not cervical carcinoma (HeLa) antigens ( C ), and reveal the frequent existence of oversized isoforms in SCA and PDAC (band 3, C–D ). GAPDH was used as the loading control. HiMark (Invitrogen) was used as the molecular weight marker. ( E ) Western blot and ( F ) FACS analyses of CSPG4 siRNA-transfected Panc1 cells further confirmed the specificity of the used antibodies and established the knockdown efficacy of the two siRNA sets (si1 and si2) at approximately 75% after 48 h post transfection, compared to the negative control siRNA (neg.si). QRT-PCR confirmed the efficacy of the knockdowns at the mRNA level.

Article Snippet: The USCN/Cloud-Clone kit employed a recombinant N-terminal fragment of a protein (Leu16-Ser350) as an ELISA standard and immunogen to produce mouse monoclonal (capture) and goat polyclonal (detection) antibodies; CUSABIO - a full-length protein expressed in eukaryotic cells.

Techniques: Western Blot, Generated, Recombinant, Derivative Assay, Control, Molecular Weight, Marker, Transfection, Knockdown, Negative Control, Quantitative RT-PCR

Journal: Oncology Letters

Article Title: High S phase kinase-associated protein 2 expression is a potential prognostic biomarker for glioma

doi: 10.3892/ol.2020.11818

Figure Lengend Snippet: Correlation of SKP2 expression with other biomarkers in 395 glioma tissues. (A) Blue and brown staining represents negative and positive expression of p-Rb, respectively. (B) According to linear correlation analysis, SKP2 was positively correlated with p-Rb in both HGG and LGG groups. ‘r’ means the coefficient of correlation. (C) Blue and brown staining represents negative and positive expression of EGFR, respectively. (D) Positive immunostaining was regarded as high EGFR expression. And no immunostaining was regarded as low EGFR expression. Mann-Whitney U test was used to evaluate the association between SKP2 and EGFR. SKP2, S phase kinase-associated protein 2; EGFR, epidermal growth factor receptor; p-Rb, phosphorylated retinoblastoma protein; HGG, high-grade glioma; LGG, low-grade glioma.

Article Snippet: The primary SKP2 antibody (1:100; cat. no. 2652S; CST Biological Reagents Co., Ltd.), anti-isocitrate dehydrogenase 1 (IDH1 R132H ) antibody (working solution; cat. no. ZM0447; OriGene Technologies, Inc.), phosphorylated retinoblastoma protein (p-Rb) antibody (1:100; cat. no. 8516T, CST Biological Reagents Co., Ltd.), epidermal growth factor receptor (EGFR) antibody (working solution; cat. no. ZA-0505; OriGene Technologies, Inc.) were added and the sections were incubated at 4°C overnight.

Techniques: Expressing, Staining, Immunostaining, MANN-WHITNEY