Mouse ascites

SC = suspension culture ARB = automated roUerbotdes MA = mouse ascites. 

Antibody. Rat monoclonal antibody 34A was purified from nu/nu mouse ascites fluid as described (79). The 34A was radiolabeled with 125I using IDO-GEN (Pierce, Rockford, IL) method to a specific activity of 2 to 4 x 105 cpm/pg, and conjugated with NGPE as previously described (7). 

Fig. 9. Conjoint Liquid Chromatography (CLC). Separation of proteins from mouse ascites and isolation of monoclonal antibody IgG in one step obtained by a combination of CIM QA and CIM Protein A Disks. Conditions Separation mode CLC (first disk CIM QA, 12 x 3 mm ID, 0.34 ml second disk - CIM Protein A, 12 x 3 mm ID, 0.34 ml, inserted in monolithic column housing) Instrumentation Gradient HPLC system with extra low dead volume mixing chamber Sample Mouse ascites Injection volume 20 pL Mobile Phase Buffer A 20 mM Tris-HCl, pH 7.4 Buffer B Buffer A + 1 M NaCl Buffer C 0.1 M Acetic acid Conditions Gradient 0 - 50 % B in 50 s, 100% A for 40 s, 100% C for 30 s Flow Rate 4 ml/min Detection UV at 280 nm... 

IgG and other proteins from mouse ascites fluid Purification CLC (combination of Anion Exchange and Affinity (Protein A) disks [76]... 

Fetner has also demonstrated chromatid breaks in a human tissue-culture cell line exposed to ozone at 8 ppm for 5 min. Other tissue-culture studies include that of Sachsenmaier et who noted tetraploidy and other chromosomal abnormalities in embryonic chick fibroblasts exposed to ozone and a decrease in transplantability of mouse ascites tumor cells. In addition, Pace et demonstrated an interference by ozone with mitotic activity in two tissue-culture cell lines. More recently, Booher et al. reported that lung cells exposed in culture to ozone concentrations as low as 0.3 ppm demonstrated an inhibition in growth that was proportional to the ozone concentration. 

OKT3 (Mab) Rescue of acute renal Mouse ascites... 

Cheng, Y.C. Prusoff, W.H. Mouse ascites sarcoma 180 thymidylate kinase. General properties, kinetic analysis, and inhibition studies. Biochemistry, 12, 2612-2619 (1973)... 

CVA, Chromosomal aberrations, mouse ascites tumour cells in vivo + 340 gg/mouse Schoneich (1967)... 

Fig. 1. SDS-PAGE minigel depicting purified IgG preparations derived from human scrum (lane 2), mouse ascitic fluid (lanes 3 and 4), rabbit serum (lane 5), and sheep serum (lane 6). Samples are electrophoresed under reducing conditions and stained with Coomassie blue. All IgGs consist predominantly of two bands comprising heavy (50,000 Mr) and light (22,000 Mr) chains with no major contaminating proteins. Molecular weight markers are shown in lanes 1 and 7 and their molecular weights given on the left.

MBP is present in various mammalian sera and activates the complement system through the classical pathway. It also specifically binds to murine monoclonal IgM and, hence, can be used to purify IgM MAb from mouse ascitic fluids. The binding reaction is calcium-dependent, so that IgM can be specifically eluted with a buffer containing a calcium chelator (e.g., EDTA). 

Dialyze mouse ascitic fluid (clarified) containing IgM against 10 mM Tris-HCl, pH 7.4, containing 1.25MNaCl. 

In addition, affinity CE separation was carried out to resolve monoclonal anti-BSA and BSA in dilute mouse ascites fluid. Then the affinity constant of this antigen-antibody interaction was measured [1004]. 

Gibb, L.E. Eddy, A.A. (1972). An electrogenic sodium pump as a possible factor leading to the concentration of amino acids by mouse ascites-tumour cells with reversed sodium ion concentration gradients. Biochem. J. 129, 979-981. 

Reid, M. Eddy, A.A. (1971). Apparent metabolic regulation of the coupling between the potassium ion gradient and methionine transport in mouse ascites-tumour cells. Biochem. J. 124,951-952. 

FIGURE 21 Separation of mouse monoclonal IgM by Immunoaffinity chromatography on rat anti-jit-chain antibodies chemically immobilized on agarose beads (4.2 mg per milliliter of resin). Column 10 mm i.d.X 45 mm. Load of 5 mL of whole mouse ascitic fluid previously filtered. Buffer phosphate buffered saline, pH 7.2. Elution by lowering the pH to 2.8 with 0.2 M glycine - HCI buffer. The first peak did not contain IgM while eluted peak (arrow) contained about 7.8 mg of IgM total. 

Clezardin, P., Bougro, G., and McGregor, J. L. (1986). Tandem purification of IgM monoclonal antibodies from mouse ascites fluids by anion-exchange and gel fast protein liquid chromatography. J. Chromatogr. 354, 425-433. 

Danielsson, A., Ljunglof, A., and Lindblom, H. (1988). One-step purification of monoclonal IgG antibodies from mouse ascites. An evaluation of different adsorption techniques using high performance liquid chromatography. J. Immunol. Methods 115, 79-88. 

Ascites Production. Anti-albumin clones HSA-1 and HSA-2 were thawed, subcloned, and reassayed by ELISA on human albumin, and a rapidly growing subclone was expanded for ascites production. Forty male BALB/c mice were primed with an intraperitoneal injection of 0.5 mL pristane on day 0. On day 14, 2 X 106 cells were injected into each mouse. Ascites fluid was collected over a 1-week period and pooled. Cells and other debris were removed by centrifugation. Approximately 180 mL of ascites fluid was obtained. 

Moser H Vecchio G (1967) The production of stable steady states in mouse ascites mast cell cultures maintained in the chemostat. Experientia 23 1-10. 

Ridgeway-Osteo-Sarcoma (Mouse) Tumor weight (g) L-1210 (Mouse) Ascites (ml) SV 40 (Hamster) Tumor weight (g)... 

Tarnowski GS, Mountain IM, Stock CC, Munder PG, Weltzien HU, Westphal O. 1978. Effect of lysolecithin and analogs on mouse ascites tumors. Cancer Res. 38 339 14... 

Fig. 4. In vivo gene delivery into mouse ascites tumor cells with Bubble liposomes. S-180 cells (1x10 cells) were i.p. injected into ddY mice. After 8 days, the mice were anaesthetized, then injected with 510 p.L of pCMV-Luc (10 p.g) and Bubble liposomes (500 pg) in PBS. Ultrasound (frequency 1 MHz, duty 50% intensity 1.0 W/cm, time 1 min) was transdermally applied to the abdominal area. In another experiment, pCMV-Luc (10 pg) - Lipofectin (50 pg) or Lipofectamine 2000 (50 pg) complex was suspended in PBS (510 pL) and injected into the peritoneal cavity of mice. After 2 days, S-180 cells were recovered from the abdomens of the mice. Luciferase activity was determined, as described in Materials and Methods. Each bar represents the mean S.D. (/j=3-6). P<0.01 compared to the group treated with plasmid DNA, Bubble liposomes, ultrasound exposure or lipofection with Lipofectin or Lipofectamine 2000. LF, Lipofectin. LF2000, Lipofectamine 2000. <10 BLU/mg protein...

The potential cytotoxic activities of forty-six Strychnos alkaloids, including harmine, were tested on different cancer or normal cells cultured in vitro. At a concentration of 1-10 pg/ml, harmine showed modest activity against L 1210 cells derived from DB A/2 mouse ascites tumor and cultured HeLa cells derived from human carcinoma. The alkaloid displayed only slight activity against cultured flow 2002 cells derived from normal embryonic human lungs, and was inactive against cultured B16 melanoma cells derived from C57BL mouse melanoma [273]. 

Tishchenko G. Neosepta microporous ion-exchange membranes in dialysis desalination of immunoglobulin fraction of mouse ascitic fluids. J Membr Sci 1996 113 237-248. 

Enzymic Reactions Involving Zinc.—A dipeptidase from a mouse ascites tumour has been shown to require zinc for the catalysed cleavage of alanine-glycine. Substitution with cobalt or manganese alters the kinetic properties of the enzyme and it is similar to other mammalian dipeptidases in other respects as well univalent anions have been found to bind to a renal zinc dipeptidase in a 1 1 mole ratio, a group with a... 

The most important technique for perfusion culture methods is to separate the concentrated cells and conditioned medium from the suspended culture broth. As noted above, the separation methods chiefly used are filtration with tubular and flat membranes as well as ceramic macroporous filters. These membrane reactors can be employed for both anchorage-dependent and suspension growing cells. Static maintenance type systems are commercially available for disposable reactors, and small size unit reactors from 80 ml to 1 liter are used for continuous production of monoclonal antibodies with hybridoma cells. The maintainable cell densities are about 10 -10 cells/ ml which is essentially mouse ascites level. However, in these systems, the cell numbers cannot be counted directly because the cells adhere to membranes or hollow fibers. Therefore, the measurement of cell density must use indirect methods. Such indirect methods include the assaying of the quantities of glucose consumption and the accumulation of lactate. The parameters of scale-up have not yet been established for these static methods. 

Reid, M., Gibb, L. E., Eddy, A. A. Biochem. J. 140 (3), 383 (1974). lonophore-mediated coupling between ion fluxes and amino acid absorption in mouse ascites-tumour cells. Restoration of the physiological gradients of methionine by valinomycin in the absence of adenosine triphosphate... 

FIGURE 3.22 West Nile virus. Immunostaining of flaviviral antigens in neurons and neuronal processes in the central nervous system from an immunosuppressed patient who died of West Nile virus encephalitis. (Flavivirus-hyperimmune mouse ascitic fluid naphthol fast red substrate with hematoxylin counterstain original magnification x40.)... 

Thyroid transcription factor-1 (TTF-1) 8C7C3/1 40-kD member of NKx2 family of homeodomain transcription factors Mouse ascites Gell Marque NA HIER... 

D2-40 D2-40 Oncofetal antigen M2A Mouse ascites Signet NA HIER... 

---