mouse anti human bcl2 mab  (Bio-Rad)

Journal: Biology

Article Title: Chemical Composition, Antioxidant Capacity, and Anticancerous Effects against Human Lung Cancer Cells of a Terpenoid-Rich Fraction of Inula viscosa

doi: 10.3390/biology13090687

Figure Lengend Snippet: IVL DCM induces the apoptosis of A549 cells. A549 cells were treated with the indicated concentrations of IVL DCM for 48 h. ( A ) P53, BCL2, BAX, Caspase 3, c-Caspase 3, PARP, c-PARP protein levels as detected by immunoblotting of A549 cell lysates. β-actin was immunoblotted as a loading control ( B ). Quantification of the bands in ( A ). Bar graphs of band intensity of target proteins normalized to the intensity of the loading control β-actin expressed as fold change of the vehicle control and represented as the mean ± SEM of three independent experiments ( n = 3). The right panel of ( B ) shows the ratio of BAX/BCL2 ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Primary antibodies used were: anti-human poly-adenosine diphosphate (ADP) ribose polymerase (PARP) 46D11 rabbit mAb which can detect the full-length and cleaved forms of PARP (ref 9532 CST, dilution 1/1000), P53 rabbit mAb (ref 2527S CST, dilution 1/500), Ki67 rabbit polyclonal antibody (ref 28074-1-AP, Proteintech, Rosemont, IL, USA, dilution 1/1000), mouse anti-human B-cell lymphoma 2 (BCL2) (ref 15071S CST, dilution 1/1000), rabbit anti-Bcl-2 associated X protein (BAX) (D2E11 ref 5023 CST; dilution 1/1000), P38 MAPK polyclonal antibody (14064-1-AP, Proteintech, dilution 1/1000), phospho-p38 MAPK (Thr180/Tyr182) antibody(ref 9211 CST, dilution 1/1000), Caspase3 antibody (ref 9662 CST, 1/1000 dilution), cleaved Caspase3 (c-Caspase 3; Asp175) 5A1E rabbit mAb (ref 9664 CST, dilution 1/1000), β-actin 8H10D10 mouse mAb (ref 3760 CST, dilution 1/1000), P21 Waf1/Cip1 12D1 rabbit mAb (ref 2947 CST, 1/1000), P27 (CST, 1/1000), phospho-FAK (p-Fak Tyr 397) D20B1 rabbit mAb (ref 8556 CST, dilution 1/1000), FAK D2R2E rabbit mAb (ref 13009 CST, dilution 1/1000).

Techniques: Western Blot, Control

Journal: Nature cancer

Article Title: Integrative genomic analyses reveal mechanisms of glucocorticoid resistance in acute lymphoblastic leukemia.

doi: 10.1038/s43018-020-0037-3

Figure Lengend Snippet: Fig. 7 | Single-cell transcriptomic analysis verifies lower CELSR2 and higher BCL2 in glucocorticoid-resistant primary ALL cells. a, Clustering of single cells (n = 2 patients) based on the top 1,000 most highly expressed genes. Both patients are independent of the discovery and validation cohorts; leukemia cells from one patient are sensitive (left) and from another are resistant to prednisolone (right). Clusters annotated to show CD19+ cells; red denotes control (untreated) and blue depicts cells after treatment with prednisolone 63 µM for 96 h. b, CELSR2 expression from clustered single-cell populations of sensitive and resistant patients either without treatment or after 96 h of prednisolone. c, BCL2 expression from clustered single-cell populations of sensitive and resistant patients. d, Bar plot depicting greater proportion of sensitive leukemia cells (n = 2,427 control cells; n = 924 treated cells) killed after treatment with prednisolone for 96 h compared with resistant patient (n = 686 control cells; n = 759 treated cells; two-proportion z-test P: ****P < 0.0001). e,f, Violin plots representing kernel density of gene expression (individual points represent single cells) of CELSR2 (e) or BCL2 (f) in CD19+ leukemia cell populations comparing prednisolone treated with untreated cells in sensitive (n = 2,427 control cells; n = 924 treated cells) or resistant patients (n = 686 control cells; n = 759 treated cells; **P < 0.01).

Article Snippet: *CHIP-seq paper* mouse anti-human BCL2 (Cell Signaling; 15071) - WB, IHC, IP, Flow mouse anti-human glucocorticoid receptor (BD Biosciences; 611227)- WB and IF validated mouse anti-human GAPDH (sc-47724)-WB and IHC validated mouse anti-human BIM monoclonal (Cell Signaling;2993)- WB and IHC, IP, Flow validated mouse anti-human GR (BD Biosciences; #611227)- WB and IHC validated mouse anti-human p-JNK (Santa Cruz Biotech; sc-6254)-WB and IHC validated rabbit anti-human cJun (Cell signaling; 9165) -WB, IP, Flow,CHIP,and IHC validated rabbit anti-human NFAT1 (Cell Signaling; 4389)- WB and IHC, IP validated rabbit anti-human phospho-cJun Ser63 (Santa Cruz Biotechnology; sc-822)-WB, IP, IFC and IHC validated rabbit anti-human LaminB1 (Cell Signaling; 12586)- WB validated mouse anti-human B-actin (Sigma-Aldrich; A5441) - Binding, ChIP, Control, Dot blot, EM, ICC, IF, IHC, IHC-Float, IP, In Vitro, and WB 3,551 citations on citeAB Anti-human CD19 PerCp-Cy5.5 (eBioscience; 45-0199-42) Flow validated Anti-mouse CD45-APC, (Tonbo; 20-0451-U100) Flow validated Eukaryotic cell lines Policy information about cell lines Cell line source(s) 697-DSMZ (ACC-42); NALM6- DSMZ (ACC 128); REH- DSMZ (ACC 22); SUP-B15 ATCC (CRL-1929); RS4;11 DSMZ(ACC 508); SEM DSMZ (ACC 546) Authentication Parental cell lines were authenticated by STR profiling Mycoplasma contamination Mycoplasma testing performed quarterly in lab on all cells all cells tested negative for mycoplasma Commonly misidentified lines (See ICLAC register) No commonly misidentified cell lines were used in this study Animals and other organisms Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research Laboratory animals NOD-SCID gamma mice, Females 8-12 weeks, n=5 mice per treatment group, Total of 60 mice on study.

Techniques: Biomarker Discovery, Control, Expressing, Gene Expression

Journal: International immunology

Article Title: Effects of BCL-2 over-expression on B cells in transgenic rats and rat hybridomas.

doi: 10.1093/intimm/dxr071

Figure Lengend Snippet: Fig. 6. Hybridoma characterization. (A) Binding constants (nanomolar) of mAbs generated with splenocytes from non-transgenic littermates (LM) and transgenic line 70818 animals (Bcl2) as determined by Biacore analysis (log scale nanomolar). (B) Western blot analysis of Bcl-2 expression in hybridomas from bcl-2 transgenic animals (each line represents one hybridoma). The positive control corresponds to a lysate from human Jurkat cells. The ratio of Bcl-2/GAPDH for each hybridoma is depicted under the respective lines. (C) Concentration of rat IgG in tissue culture supernatants at day 3 of culture (cells between 3.1 and 6 3 105 ml–1) measured by ELISA. Each point represents the values of one hybridoma supernatant and the horizontal bar the mean of each group of values *P < 0.05 hybridoma Bcl-2- (n = 4) versus hybridoma Bcl-2+

Article Snippet: Western blots were performed as previously described (26) using a mouse anti-human Bcl2 mAb (Abd Serotec) followed by incubation with HRP-conjugated goat anti-mouse IgG + IgM (H + L) antibodies (Jackson ImmunoResearch).

Techniques: Binding Assay, Generated, Transgenic Assay, Western Blot, Expressing, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay