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- Goal: show A-beta binding to LilrB2/PirB-eYFP.
- Micrograph: fusion protein (yellow), nucleus (blue), antibody (red)
- A-beta has biotin conjugated; need streptavidin/red.
- Streptavidin - Alexa Fluor 594: $185 for 1 mg http://www.lifetechnologies.com/order/catalog/product/S11227
- GFP chicken polyclonal ICC/WB http://www.abcam.com/gfp-antibody-ab13970.html
- Goal: show LilrB2 and PirB localized to the membrane
- Micrograph: fusion protein (yellow) (or maybe anti-GFP ICC), nucleus (blue), ab (red)
- No LilrB2 proteins verified for ICC. Could ask for an Abreview discount on mouse polyclonal http://www.abcam.com/lilrb2-antibody-ab172538.html
- No PirB abs verified for ICC. Could ask for an Abreview discount on mouse monoclonal http://www.abcam.com/lilrb3-mm0413-9s32-antibody-ab90939.html
- Secondaries: goat/donkey anti-mouse AF594
- Goal: cofilin western blot
- mouse monoclonal ICC/WB http://www.abcam.com/cofilin-antibody-ab42824.html
- rabbit monoclonal ICC/WB (no lead time!) http://www.abcam.com/cofilin-ep6376-antibody-ab134963.html
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- Goal: show that the IgM makes it to the cell membrane
- Expected micrograph: nucleus (blue), fusion protein (yellow), antibody (red)
- Abcam rabbit anti-human IgM (Biotinylated): http://www.abcam.com/rabbit-human-igm-mu-chain-biotin-ab97208.html ($165)
- Biotin detection may not be the best way forward: the reason that immunihistochemistry is so sensitive is the amplification effect of a good secondary.
- May be better to get a rabbit anti-human IgM and then use a donkey anti-rabbit 594 secondary
- Abcam has validated this mouse anti-human IgM (Fc fragment) for ICC, flow, WB:
- http://www.abcam.com/human-igm-fc-fragment-ch2-antibody-ab772.html (will order this one, use PG's anti-mouse AF594 secondary and the 800CW IRDye for WBs)
- Want mu-chain binding to help experimentally activate the BCR (hence the secondary.) Will get a conjugated secondary for ICC too.
- Goal: Syk for western blot
- Found secondaries! 800-CW goat anti-mouse, 680RD (or LT) goat anti-rabbit
- Li-Cor recommends a rabbit primary for the low-abundance protein.
- Rab-mab validated for WB, ICC: http://www.abcam.com/syk-ep573y-antibody-ab40781.html
- Secondary: 680RD anti-rabbit IRDye
- Found secondaries! 800-CW goat anti-mouse, 680RD (or LT) goat anti-rabbit
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