Anti CASZ1  

Western blot using Rockland's anti-hCASZ1 antibody. This blot shows detection of endogenous and transfected human CASZ1 protein in fresh whole cell lysate (~30 μg). Lane 1: BE2(s) cell lysate. Lane 2: BE2(N) cell lysate. Lane 3: SY5Y transfected with hCASZ5 (125kDa). Lane 4: SY5Y transfected with hCASZ11 (190kDa). Protein was resolved by SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1,000 for 1.5 hours at room temperature then incubated with HRP-conjugated Goat Anti-Rabbit antibody for 45 min. at room temperature. Personal communication, Carol Thiele, NCI, Bethesda, MD.

Western Blot of Anti-CASZ1 Antibody. Lane 1: NBLS Cytoplasmic (20μg). Lane 2: NBLS Nuclear (3μg). Lane 3: BE2C Cytoplasmic (30μg). Lane 4: BE2C Nuclear (7μg). Lane 5: SY5Y-CASZ1b (10μg). Block: 5% Blotto/TTBS for 1 hour. Primary: Casz1 1:10,000 for 1 hour. Secondary: Goat anti-Rabbit HRP for 1 hour. 240sec exposure. Detects nuclear endogenous CASZ1a and CASZ1b; and transiently transfected CASZ1b isoform. Personal communication and images from Carol Thiele Galetto, NCI.

(A) a. CASZ1 localized exclusively in the cytoplasm showed by staining the tumor samples with anti-CASZ1 antibody; b. CASZ1 localized to both the nucleus and the cytoplasm. Nucleus was counterstained with hematoxylin solution showed in blue, and CASZ1 was probed with rabbit anti-CASZ1 antibody and showed in brown. Fig 9. PMID:？27270431

(B) Examples of nuclear staining: a. Score 0 - a rare positive nuclei; b. Score 1 (1？10% positive) equivocal/uninterpretable; c. Score 2 (10？50%) weak positive, or d. Score 3 (> 50% positive cells)-strong positive. Fig 9. PMID:？27270431

(A) Endogenous CASZ1 subcellular localization in BE2 cells was determined by staining the cells with rabbit anti-CASZ1 antibody. The anti-CASZ1 antibody was pre-incubated with (bottom panel) or without (top panel) CASZ1 antigen peptide (Pep) that used to generate CASZ1 antibody. The chromatin was stained with DAPI. Fig 8. PMID:？27270431

(B) Mouse xenograft tumor of human NB cell line that has been stably transfected with tetracycline inducible CASZ1 (NGPtetCASZ1) was dissected and stained with rabbit anti-CASZ1 antibody and DAPI. Fig 8. PMID:？27270431

(C) Endogenous CASZ1 in the eye of adult mouse was stained with rabbit anti-CASZ1 antibody and DAPI. Localization: nucleus in lens epithelia but primarily localizes in the cytoplasm in photoreceptor cells. Fig 8. PMID:？27270431

A. Anti-CASZ1 antibody is specific to CASZ1a and CASZ1b. Induction of CASZ1a and CASZ1b expression in the SY5YtetCASZ1a and SY5YtetCASZ1b clones by Tet at 24 hours was visualized by immunoblotting the cell lysates with anti-CASZ1 antibody (left); endogenous CASZ1a and CASZ1b expression in neuroblastoma cells was visualized by immunoblotting the BE2 cell lysates with anti-CASZ1 antibody, and the specificity of the antibody was demonstrated by pre-incubation of immunizing peptide and loss of CASZ protein recognition with immunoblotting analysis (right). B. Endogenous CASZ1a and CASZ1b are predominantly expressed in the nucleus of neuroblastoma cells. The nuclear localization of CASZ1a and CASZ1b was characterized by the co-localization of Alex 568-labeled goat anti-mouse IgG and DAPI-stained nucleus of BE2 cells (top), and the specificity of the antibody was demonstrated by the blocking effect of antigen peptide on antibody recognition with immunoblotting analysis (bottom). C. The co-expression of murine CASZ1a and CASZ1b protein in heart, lung and eye of P6 mouse was visualized by immunoblot analysis of tissue lysates using an anti-CASZ1 antibody. D. The simultaneous up-regulation of CASZ1a and CASZ1b protein by either RA or 5-Aza-dC in neuroblastoma cell lines was visualized by immunoblot analysis of the cell lysates using the anti-CASZ1 antibody. Fig 3. PMID:？21490919

(B) Western blot analysis of specific associated polypeptides using elutions from TAPs.？Fig 1. PMID:？23592795

d？ChIP-seq was performed using anti-CASZ1 antibody in SMS-CTR cells with or without MEKi treatment. Composite plot shows the increase of endogenous CASZ1 binding on genomic DNA after SMS-CTR cells are treated with 100？nM MEKi for 48？h. Fig 2. PMID:？32060262