http://www.protocol-online.org/forums/index.php Fri, 20 Nov 2009 15:47:40 -0800 360 http://www.protocol-online.org/forums/index.php?showtopic=11658 Wed, 18 Nov 2009 18:10:47 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11658 http://www.protocol-online.org/forums/index.php?showtopic=11633 ]]> Wed, 18 Nov 2009 00:57:18 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11633 http://www.protocol-online.org/forums/index.php?showtopic=11618 Hi everybody. After purification of a membrane fraction, I have measured the absorbance at 215nm (peptide bond absorption) and at 280nm (absorption by aromatic amino acids (tyrosine, tryptophan and phenylalanine)). Now, at 215 nm the peak corresponding to the monomer is sharper than at 280 and the peak corresponding to the aggregates almost disappears. THe preparation is identical in both measurements. Any hint on why this is happening? And which measurement at this point is most reliable? Thanks a lot! Tue, 17 Nov 2009 13:15:34 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11618 http://www.protocol-online.org/forums/index.php?showtopic=11592
Can someone please tell me what ALL are the scientific reasons when Immunofluorescence staining (in cells) is positive and Western blotting negative.
Ofcourse what I see in immunofluorescence staining is positive and not any unspecific staining. shRNA knockdown has confirmed this.
where i can read the scientific reasons for this case.
]]> Mon, 16 Nov 2009 18:27:51 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11592 http://www.protocol-online.org/forums/index.php?showtopic=11471 Wed, 11 Nov 2009 11:33:29 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11471 http://www.protocol-online.org/forums/index.php?showtopic=11461 I am trying to make a 10mls of ATP 10Mm solution the MW is 605.2

My balance is in grams and I am struggling...
any help would be good.

]]> Wed, 11 Nov 2009 04:36:40 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11461 http://www.protocol-online.org/forums/index.php?showtopic=11351 We are following the USP Typsin Activity Assay. Trypsin is suppose to convert the substrate BAEE + H2O to Na-Benzoyl-L-Arginine + Ethanol and the change should be seen via UV Spec at 253nm at a rate of 0.003absorbance units/min. We are seeing no change at all. Anyone have any ideas why this change might not be taking place? We are using a Sigma Trypsin reconsituted in 0.001N HCL. Thanks.]]> Wed, 04 Nov 2009 09:05:57 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11351 http://www.protocol-online.org/forums/index.php?showtopic=11321

i've been working with a mitochondrial carrier with aims to reconstitute it into liposomes. After the protein has been expressed in bacteria, I solubilize it with ~3% TX-100. The recontitution takes place mixing protein, PC, cardiolipin, TX-114 & internal susbtrate to further load the mixture (x24) into an Amberlite column (anion exchanger).

Excess of external substrate is removed using a Sephadex G75. The transport assay is performed after this.


So far I have been unable to observe transport using this conditions. Does anyone have any experience/comments/advice for my issue


Thanks


P.]]> Tue, 03 Nov 2009 07:32:53 -0800 http://www.protocol-online.org/forums/index.php?showtopic=11321 http://www.protocol-online.org/forums/index.php?showtopic=11113 HEy there.. Can anyone tell the the change in pH (interms of phnit/sialic acid) of a glycoprotein on addition of a single sialic acid?!! Mon, 26 Oct 2009 00:03:46 -0700 http://www.protocol-online.org/forums/index.php?showtopic=11113 http://www.protocol-online.org/forums/index.php?showtopic=11060
i am dealing on isolation of antimicrobial bioactive compounds from plant. however, through out my study i found that responsible bioactive compounds from the bioactive fraction always different in different batch of sample collection. so it is so difficult to hv a consistant bioactive supply. Anyone of you hving experiences on these and how to mangange this kind of problem? what the variable that should be taken more in order to hv repeatabe bioactive pattern in the bioactive fraction?

thank for your help guys~~~]]> Fri, 23 Oct 2009 00:34:23 -0700 http://www.protocol-online.org/forums/index.php?showtopic=11060