**Note this folio is actual from John Tullai and if you accept a articulation botheration try here
In this section, the answers are either placed in the anatomy of this page, or, area applicable, a articulation to download a book (usually a pdf) is provided.
The action and discharge filters are audible for all the dyes and Sypro ruby. The above botheration is that Sypro bittersweet fluoresces beyond such a advanced ambit (excite about 400, discharge about 630) that you will get some Sypro bittersweet arresting if you were to try to re-image the Cy dyes. This agency that already you’ve decrepit with Sypro ruby, you can no best go aback and re-acquire a Cy dye image, back the quantitation would no best be correct. So there is cross-talk in the Cy dye channels due to arresting advancing from Sypro ruby.
For low alkali digestion, use 7-20mM ammonium bicarbonate/50% booze instead of 50mM ammonium bicarbonate/50% booze solution.
Cy Dye apprenticed to the peptides and altitude with Accumulation Spec:
The basal band is that due to the actual low affluence of the Cy Dye to be potentially apprenticed to the protein that it is not an affair in apprehension with the basal labeling (to the lysine residues) (the assimilation labeling through the cysteine balance is accumulation spec. accordant and needs the Cy Dye MWt to be added into the abstracts abject chase for actuality apprenticed to the cysteine residues)
Immobilized pH gradients-table of gradients. Download the pdf (1 MB)
Sypro Bittersweet staining agreement of 2D gels. Download the pdf (68K)
The capricious stain for autumn IPG strips at allowance temp. Download the pdf (12K)
Bind-silane adjustment for gelbond-backing gels. Download the pdf (8K)
Reference anecdotic the adjustment to accomplish new Sypro Bittersweet stain. Pubmed Reference.
Reference for active IEF of proteins with basal pI. Pubmed Reference
Basic pH acclivity with aerial sample loads. Pubmed Reference. Brief adjustment pdf (4K)
Reference anecdotic access in [DTT] to 3% to advance basal pI resolution. Pubmed Reference.
Acid violet staining of immmobiline gels and strips: Patesotos et al. (1988) Electrophoresis 9:488-496. Download the pdf of the agreement (4K).
The use of glycerol and isopropanol in the rehydration absorber and altercation of electroendosmosis. Download the pdf
Test sample protocols-trial preps that about consistently assignment able-bodied for 2DE:
The use of IPG band with berserk proteins. Pubmed Reference.
Capacity of polyacrylamide gels. Pubmed Reference.
Diffusion Blot protocol-western adjustment acclimated for backed gels-gels can again be added candy (Spot picker, etc.). Download the pdf (4K).
Spot-picker ambit for ailing gels. Download the pdf (4K).
The Tris tricine absorber arrangement for the DALT 12. Download the pdf (4K)
Molecular Wt. against gel allotment – Look at folio 635 of the Amersham 2002 catalog-the table is at the bottom
The kits for film apprenticed protein abstraction etc is here: http://www.genotech.com/
Cell Ablution for able beef above-mentioned to agriculture for IEF:
Salt from advance average or ablution band-aid can be significant. Salt-free buffer/osmoticum should be acclimated for abrasion (10 mM Tris / 250 mM sucrose pH 7.0)
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