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HELP! LPS stimulated plasma gone awry [View Printable]
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Nimoy
Group: Member
Posts: 8
Joined: Oct 12, 2005
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I have cultured rat blood and stimulated it with LPS... I followed a protocol I found in the literature, however, the levels of cytokines produced are at the lower limit of detection with my R+D systems TNF ELISA kit.... The levels in published are at least 20 times what I am finding.... Can anyone tell me how my cultures have gone wrong? |
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| Posted Nov 08, 2005, 8:05 AM |
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RockyDoc
Group: Member
Posts: 20
Joined: May 02, 2006
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Nimoy,
Did you find out what happened with your LPS-stimulated cells? I am currently working with other stimulants (Concanavalin A, PMA/IoM) and I am not getting the results reported in the literature. Just wondering whether I may be going through what you experienced?
Kind regards,
RockyDoc
| Nimoy said: | | I have cultured rat blood and stimulated it with LPS... I followed a protocol I found in the literature, however, the levels of cytokines produced are at the lower limit of detection with my R+D systems TNF ELISA kit.... The levels in published are at least 20 times what I am finding.... Can anyone tell me how my cultures have gone wrong? |
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| Posted Aug 28, 2006, 10:17 AM |
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Nimoy
Group: Member
Posts: 8
Joined: Oct 12, 2005
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| RockyDoc said: | Nimoy,
Did you find out what happened with your LPS-stimulated cells? I am currently working with other stimulants (Concanavalin A, PMA/IoM) and I am not getting the results reported in the literature. Just wondering whether I may be going through what you experienced?
Kind regards,
RockyDoc
| Nimoy said: | | I have cultured rat blood and stimulated it with LPS... I followed a protocol I found in the literature, however, the levels of cytokines produced are at the lower limit of detection with my R+D systems TNF ELISA kit.... The levels in published are at least 20 times what I am finding.... Can anyone tell me how my cultures have gone wrong? |
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| Posted Aug 28, 2006, 11:40 AM |
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Nimoy
Group: Member
Posts: 8
Joined: Oct 12, 2005
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Yeah, I did find out what the problem was.. I had used an EDTA coated tube, which was apparently suppressing my stimulated cytokine responses... I have now switched to sodium heparin coated tubes and the levels are much higher... Hope this helps!
| Nimoy said: | | RockyDoc said: | Nimoy,
Did you find out what happened with your LPS-stimulated cells? I am currently working with other stimulants (Concanavalin A, PMA/IoM) and I am not getting the results reported in the literature. Just wondering whether I may be going through what you experienced?
Kind regards,
RockyDoc
| Nimoy said: | | I have cultured rat blood and stimulated it with LPS... I followed a protocol I found in the literature, however, the levels of cytokines produced are at the lower limit of detection with my R+D systems TNF ELISA kit.... The levels in published are at least 20 times what I am finding.... Can anyone tell me how my cultures have gone wrong? |
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| Posted Aug 28, 2006, 11:43 AM |
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