Recommendation years agoKeWei ZhaoVA Greater Los Angeles Healthcare SystemA bit of historical background in Western Blotting involving antiphosphate antibodies please forgive if you knew the story old days only tyrosine Pserine threonine Abs were available and blocking agents such milk which contains Pproteins had be avoided. Unless there a different instruction from antibody manufacturer minor changes can be made according to applications such as fluorescent IHC rule is use milk for Western Blots and BSA . years agoRaj KumarUniversity of Texas MD Anderson Cancer CenterMost the phosphoprotein antibody kit suggest to use BSA blocking agent but some time phosphoproetin also work using milk you try by both [...]

Cyril and Methodius UniversityIf you need to detect phosphoproteins pAkt pErk etc ALWAYS use BSA TBS for blocking. So Dr [...]

BSA CST in TBS for blocking antibody incubation is the best start primary most influential outcome quality. Answers years agoSandeep University in St. unbind opfOpenEnd w sj evt re opfOpenStart else function be var et chromewebstore item chromeinline extn ef ft ot ge opalpers anch flyout onP appHTML if ildNodes moveChild for CI yleExp sendBeacon navigator fd lsp px log return setHeight [...]

Length do if ift r art break . So Dr. While it sounds reasonable to use BSA for blocking sometimes you might get high background due insufficient . A is thought to work better for phospho antibodies as albumin secreted protein and tends not phosphorylated [...]

In my hands milk works best for me all the antibodies. When we had to perform difficult phosphorylated Smad IHC ve used basic protocol with PBST and the ABC kit. in var c body e h if return setTimeout q . answers addedHow to store membrane in Western BlotI am beginner [...]

Recommendation years agoDmitry OvchinnikovThe University of fist bet WBdedicated BSA from CST albeit expensive Tween used . generally in the case of phosphopreotin we use higher amount protein. for example pSK T clone D from Cell Signal is cleaner if the membrane blocked in milk rinsed few times then probed BSA. Recommendation years agoMd Wasim KhanUniversity of Illinois ChicagoMilk works fine for [...]

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PLEASE let me know and correct it if unconsciously forget mention the authors who give answers included below. You just have to try your antibodies and see what works best. years agoFrancesca PortsmouthI have been working with phosphoproteins as well. months agoBoris AleksovskiSs [...]

And our Bing your search engineGet smarter earn rewards faster by adding the New Tab with extension. rights and is one of the most important biomass resources. a year VenkatesanTamil Nadu Veterinary and Animal Sciences UniversityI used BSA TBS to check phosphorylation of proteins results were good. If m not mistaken we ve incubated the samples with primary antibody overnight [...]

In my experience TBST is good buffer. and our Bing your search engineGet smarter earn rewards faster by adding the New Tab with extension. Recommendation years agoAlexandre Gon alvesUniversity of ZurichHi Diptadip Dattaroy Can you please check the figure my paper Reduction BMP activity by gremlin enables ureteric bud outgrowth GDNF WNT feedback signalling during kidney branching morphogenesis [...]

However insufficient blocking might occur as former excellent answers mentioned. Recommendationa year agoMushtaq Ahmad AnsariKing Saud Universityi use both BSA and milk TBST buffer results are very good. Has anyone found that TBS inhibits their western blots is couple of years old those cubes x would this likely affect chemistry concentrations etcIf not transfer issue something else do my head agoRongqing Aaron PanAdded answerFor WB with phosphoprotein think most important to dilute your primary antibody BSA solution might overwhelmed and sequestered by milk Casein [...]

Loss of blotted proteins A study demonstrated that soaking membrane milk solution can cause from the . V [...]