Amal Halabi added an answer:What can I do when newly purchased RAW 264.7 does not respond to LPS (10-500 ng/mL)?We recently purchased macrophage like cell RAW 264.7 from European collection of cell cultures. The cells have been thawed according to original handling procedure for frozen cells. We have now passage no.10. They seems completely healthy, no sign of contamination, no visible morphological changes. Unfortunately, the cells do not respond to LPS (10-500 ng/mL) at all, measured with Griess reagent as NO production. Griess reagent is new and fresh. The positive control (LPS) does not differ from negative control +/- 4 uM NO. LPS was previously tested on "old" RAW cells and it worked even at low concentration 10 ng LPS/mL.
Because these cells are new and I want to avoid to buy new one, I kindly ask you for some suggestion what can I do? Should I try to apply CSF1 (M-CSF) or any other stimulus to stimulate them to start respond? Or does exist some other easier solution?Hello Ondrej
iF you still have the old RAW ,you can try activate your new one by adiing 1:1 old/new medium (old medium = supernatant mediium from the old growing RAW). If this did not work try to purchase from ATCC.,Good luck.Following
Bhaskar Vemu added an answer:What is the difference between lead nitrate and lead chloride bioavailability in sheep or in another species after repeated oral administration?I am conducting an experimental design in view to relate between repeated lead nitrate oral administration to non-lactating ewes with toxicological responses such as oxidative stress and immunotoxicity (Immuglobulins, B and T Lymphocyts). I am surprised to find a value of blood lead lower than that observed after repeated lead chloride. Note that the accuracy of analytical method is good.has anyone tried to test the in vitro absorption assessment using caco cells. Also it is true that different formulations have different absorption capacities, but, the volume of the liquids administered orally, concentration of lead salts in the solution and the feeding status (i.e., fasting/post prandial) should also be seen. As we all know that rumen contains lot many living organsims, we should also see if the alteration in pH because of chloride or nitrate or nitrite or ammonia affects the absorption of lead (due to complex formation). we should see if lead absorption follows a zero or first order kinetics as well.Following
Reza Banihashemi added an answer:Which bacterial toxin is better for the treatment of cancer?Immunotoxin therapy.Hi thanks a lot this toxine is very general by several problemFollowing
Marina Ninkov asked a question:Can anyone help me with lymphocyte isolation from a rat's small intestine?Is it possible to use 2-mercaptoethanol instead of DTT?Following
Ioannis D Kyriazis added an answer:Can any one suggest a method to study macrophage activity in fish head kidney?Head kidney is the main immunological organ in fishesYes. You could check the phagocytosis ability of and capacity of isolated macrophages from Head Kidney tissue...You can check it with yeasts or with fluorescent beads through FACS analysis.
You can check the below paper that is published by the laboratory that I worked. These days we participated in a new programme and i advanced this technique with fluorescent beads through FACS Analysis...if you search more in pubmed you can find many procedures with trivial alterations..
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Karagouni E, Athanassopoulou F, Tsagozis P, Ralli E, Moustakareas T, Lytra K, Dotsika E.
Int J Immunopathol Pharmacol. 2005 Jan-Mar;18(1):121-32.
The impact of a successful anti-myxosporean treatment on the phagocyte functions of juvenile and adult Sparus aurata L.Following
Study of immune response to xenbiotic and foreign substance exposure.