monoclonal rabbit anti-human ki67/mki67  (Novus Biologicals)

Journal: Cell Biology International

Article Title: FLOT1 Is a Novel Serum Biomarker of Ovarian Cancer Targeted by N6‐methyladenosine Modification Inhibition

doi: 10.1002/cbin.70015

Figure Lengend Snippet: Effect of m 6 A modification on FLOT1 expression and tumor formation. (A) Heatmap showing unsupervised clustering for 26 genes of m 6 A RNA modification in OC patients ( n = 376). Each column represents patients, and each row represents an m 6 A RNA modification regulator. (B) Heatmap of the correlation coefficient for the interaction between FLOT1 and the m 6 A RNA modification regulator. Red indicates a positive correlation, and green indicates a negative correlation. (C) Detection of FLOT1 mRNA expression in two human ovarian cancer cell lines (OVCAR‐3 and A2780) and a normal human ovarian surface epithelial cell line (IOSE‐80) by RT‐qPCR. Two‑way ANOVA followed by the Sidak test was used. The level of FLOT1 mRNA expression was higher in OVCAR‐3 and A2780 cells than in IOSE‐80 cells ( n = 3). (D) The levels of total m 6 A methylated RNAs in OC cells were verified by colorimetrically quantified ( n = 3). (E) Detection of the m 6 A modification level of FLOT1 mRNA in IOSE‐80, OVCAR‐3, and A2780. (F) Prediction of FLOT1 m 6 A modification sites by SRAMP. (G) Measurement of FLOT1 mRNA expression in OVCAR‐3 and A2780 cells detected by RT‐qPCR after 3‐deazaadenosine (3‐DAA) treatment. A Student t ‐test was used ( n = 3). (H) Detection of the m 6 A modification level of FLOT1 mRNA by MeRIP‑qPCR after 3‐DAA treatment. A Student t ‐test was used ( n = 3). (I) The workflow of ovarian cancer xenograft model construction and 3‐DAA treatment. (J) Xenograft tumor formation in nude mice. OVCAR‐3 cells were subcutaneously implanted in nude mice ( n = 5/group). Animals were photographed on day 30. Measurement of mouse weight. (K) Photo of executed tumors. Live measurement of tumor volume. (L) Images of the H&E and immunohistochemistry staining of FLOT1 and Ki67. Original magnification x200; Scale bar, 200 µm. Partial magnification x600; Scale bar, 50 µm. (M) IHC staining score (ISS) of FLOT1 and Ki67 protein expression. (N) Evaluation of FLOT1 protein expression by Western blot and semi‐quantification of the blot in the 3‐DAA treated (+) and untreated (−) groups. The data are shown in the form of the mean ± SD. A Student t ‐test was used. ns, not sigificance; * p < 0.05; ** p < 0.01; **** p < 0.0001.

Article Snippet: Co. Ltd., Fuzhou, Fujian, China) for 40 min at room temperature, the sections were incubated with a primary rabbit monoclonal anti‐human FLOT1 antibody (1:100 dilution; RRID: AB_11156367, Cat# ab133497, Abcam, Melbourne, Australia) at 4°C overnight, followed by a HRP‐conjugated anti‐rabbit secondary antibody (1:500 dilution; RRID: AB_2811189; Cat# GB23303, Servicebio, Wuhan, China) for 1 h at room temperature or a rabbit monoclonal anti‐human Ki67 antibody (1:100 dilution; RRID: AB_2834152; Cat# AF0198, Affinity Biosciences, OH, United States).

Techniques: Modification, Expressing, RNA modification, Quantitative RT-PCR, Methylation, Immunohistochemistry, Staining, Western Blot

Journal: Heliyon

Article Title: Bioinformatics in vivo and in vitro assays identified miR-486-5p as a tumor suppressor miRNA in hepatocellular carcinoma

doi: 10.1016/j.heliyon.2024.e39909

Figure Lengend Snippet: miR-486-5p overexpression inhibited HCC growth in vivo . (A, B) Tumor fluorescence intensity detected using in vivo bioluminescence imaging in nude mice injected with HepG2 cells transfected with miR-486-5p lentivirus expressing luciferase under the liver envelope. (C) Representative morphological and HE-stained HCC sections. (D) miR-486-5p expression in HCC tissues in the NC group and the miR-486-5p group. (E) IHC staining of PCNA in HCC tissues in the NC and miR-486-5p groups. (F) IHC staining of Ki67 in the HCC tissues in the NC and miR-486-5p groups. ∗∗∗p < 0.001.

Article Snippet: The sections were then incubated overnight with primary antibodies, either mouse anti-human PCNA antibody (ab29, Abcam) or rabbit anti-human Ki67 antibody (27309-1-AP, Proteintech), at 4 °C.

Techniques: Over Expression, In Vivo, Fluorescence, Imaging, Injection, Transfection, Expressing, Luciferase, Staining, Immunohistochemistry

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of a Myeloid Cell Leukemia 1 (Mcl-1) Inhibitor That Demonstrates Potent In Vivo Activities in Mouse Models of Hematological and Solid Tumors

doi: 10.1021/acs.jmedchem.4c01188

Figure Lengend Snippet: Efficacy study of 26 in the A427 cell line-derived NSCLC xenograft model. A. Cell viability measurement following treatment with 26 (left) or docetaxel (right) at indicated doses. Viability is normalized to control population measurement at the end of the treatment duration. B. Heatmap of cell growth inhibition. C. Synergy between the two treatments based on BLISS Gap score. D. Tumor volume of control (gray), 26 dosed at 60 mg/kg q7d (orange), docetaxel dosed at 10 mg/kg q7d (teal), and 26 dosed at 60 mg/kg q7d + docetaxel dosed at 10 mg/kg q7d (beige). E. Average body weight for each dosing cohort: control (gray), 26 dosed at 60 mg/kg q7d (orange), docetaxel dosed at 10 mg/kg q7d (teal), and 26 dosed at 60 mg/kg q7d + docetaxel dosed at 10 mg/kg q7d (beige). F. Comparison of relative tumor volumes for each dosing cohort. G. Tabulation of dose, AUC, % TGI, and % tumor regression. H. Representative images of hematoxylin and eosin (H&E) and Ki67 stained tumor sections for each dosing cohort. I. Summary of tumor viable area in mm 2 and percentage of Ki67 positive cells for each tumor and treatment group.

Article Snippet: Ki67 staining was performed on the automated platform BOND RX (Leica) using a rabbit antibody against human Ki67 (clone D2H10 from CST) at 1:400 dilution with 20 min heat mediated antigen retrieval at pH 6, followed by DAB.

Techniques: Derivative Assay, Control, Inhibition, Comparison, Staining

Journal: Cell Death & Disease

Article Title: Human papillomavirus-associated head and neck squamous cell carcinoma cells lose viability during triggered myocyte lineage differentiation

doi: 10.1038/s41419-024-06867-4

Figure Lengend Snippet: A Xenograft tumor of P1 with cystic/necrotic core. Hematoxylin/eosin (HE) and p16 staining of the same paraffin block were shifted into a similar orientation for comparison. B IF staining of P1 xenograft tumor reveals cells co-expressing TPM1 and KRT17 (arrow) and tissue positive for TPM1 and EpCam (star); 10x magnification. C P1 xenograft tumor and P3 original tumor show Ki67 in areas with lower TPM1/KRT17 expression; scale bars = 10 µm.

Article Snippet: Primary antibodies: rabbit-anti-human ACTA1 (1:200, EPR16769, ab179467, Abcam, Cambridge, UK), rabbit-anti-human CDKN2A/p16INK4a (EPR1473; 1:200; Abcam), rabbit-anti-human EpCam (EGP40/1556R; 1:100; Novus Biologicals, Centennial, CO, United States), mouse HPV antibody cocktail (1:100, CAMVR-1 & C1P5, Z2657MS, Thermo Fisher Scientific, Waltham, MA, United States), rabbit-anti-human Ki67 (1:200, SP6, ab16667, Abcam), mouse anti-human Tropomyosin (1:100, F-6, sc-74480, Santa Cruz Biotechnology, Dallas, TX, United States).

Techniques: Staining, Blocking Assay, Comparison, Expressing