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Awfulinvestor, my apologies. I was looking at Poirot's link (www.sigmaaldrich.com/ES/es/product/mm/mabs1918) and I read the figure wrongly. In the EU they use a comma to separate Euros from cents. I saw the comma and my English brain computed 'thousand'. I should know better.
Thanks for the 'ug' explanation too. As a non-scientist I don't completely get the technical explanations, like yours, which appear on here, but I learn a little more every time I read them. And they help to 'colour in' more bits of the emerging bigger picture which I think is looking pretty good.
Apre: Where did you get £120,000 Euros from? It’s £113 for 25ug.
And that is quite a lot of material. You have to take into account the size of the protein (affimer) you are buying. Affimers are small (compared to an antibody) at 16kDa (IgGs are about 150kDa), so when working out the molarity, you take this into account. Ie in 25ug of either affimer or IgG, the number of molecules of affimer will be 10x higher (approx) than the IgG.
E.g. if you bought that 25ug, you’d either receive it as a liquid or as a tiny spec of powder in a tube (in this case you’d resuspend it in water, as the buffer contains pbs). I can’t tell without a CoA how they give it to you, it says ships on dry ice, so I guess liquid that is frozen and then the first time you use it you thaw and aliquot smaller amounts.
Say they give to you at 100ul. That is 250ug/mL. The size is 16kDa.
(My brain normally hurts doing these calculations. I just remember 1mg/ml for IgG is /0.15 (150kDa) = 6.66uM, or if you divide by 150,000 it’s 0.0000067M.)
So for this: 250ug/ml = 0.25mg/ml
0.25/0.016 (16kDa) = 15.6uM.
As the affinity (kD) is 20pM (very high affinity), you are way above that at 15.6uM. (About 1 million fold). That’s why it says do a 1:1000 dilution, even this is probably wasting a lot of your material!
Hope this makes some sense. TLDR: a ug of something is not always tiny!
That article was dated 05 Oct 2017.
Towards the end of the link for the published papers I just posted it says:
'Contact for Reagent and Resource Sharing
Requests for further information or reagents should be directed to the lead contact and corresponding author, David Komander ( dk@mrc-lmb.cam.ac.uk ). Request for cell lines from other studies should be directed to the respective corresponding authors. Affimers can be obtained from Avacta (https://www.avacta.com).'
Thanks Matml74, that's very helpful.
Been around since 2017 @yenom so any revenue generated from these reagents will be recored in the annual accounts.
https://avacta.com/2017-09-25/
Top notch sleuthing there Poirot. Thank you.
Just an observation from a very definite non-scientist who can google a bit.
So a "ug" is a microgram, or one millionth of a gram, or one thousand of a milligram. And they are selling 25ug for 120,000 Euros. Now I don't know how many grams of reagent a lab may need for whatever process . . . but it strikes me (and scientists please step in here) that if you are working on large scale projects then that 120K Euros per unit could multiply pretty quickly.
In revenue terms, selling a relatively small quantity of reagent is potentially somewhat more lucrative than selling umpteen thousand LFT's . . . and we look like we are going to be selling somewhat more than umpteen tousand of them.
I've said it before and I'll say it again, the bigger picture just keeps getting bigger and bigger . . . the mind bugles. (That's boggles but using micrograms . . . see what I did there?)
Should there have been an RNS covering this?
Research. Other biotech/academia/pharma.
Who and for what use are these affimers being purchased?
Avacta’s Japanese distributor has them for sale also, along with more of the catalogue Affimers.
https://www.cosmobio.co.jp/product/detail/affimer-product-als.asp?entry_id=14177
Good to see Affimers getting commercial traction now, clearly the exposure of the last 12 months is helping massively in that regard:)
Hi Timster Does refer to Affimer® ( Avacta trademark ) in Merck Product Information.
"Description Anti-Ubiquitin Lys6 specific Affimer® reagent GFP/His tag" etc
Presentation Purified GFP/His-tag Affimer® reagent in buffer containing 100 mM Sodium Phosphate, 150 mM Sodium Chloride, 0.02% Sodium Azide, pH 7.4.
Quality Level MQ200
Applications
Application Anti-Ubiquitin Lys6 specific GFP/His-tag Affimer® reagent, Cat. No. MABS1918, detects diubiquitin K6-linkage and is tested for use in Western Blotting.
Key Applications
Western Blotting
Application Notes Western Blotting Analysis: A representative lot of this Affimer® detected di-Ubiquitin Lys6. in Western Blotting applications (Michel, M.A., et. al. (2017). Mol Cell. 68(1):233-246)."
This might be the Avacta product sheet ?
Product Name Anti-diUbiquitin K6-linkage Affimer (44-29)
Catalogue Code AVA00015
Description Affimer (44-29) to diUbiquitin K6-linkage
Clone ID 44-29
Tested Applications Direct ELISA
Tags C-term 6His
Conjugate Non
https://www.drugtargetreview.com/wp-content/uploads/Affimer-44-29.pdf
General description
Ubiquitin (Ub), a highly conserved 76-amino acid (8.6 kDa) protein that escorts proteins for rapid degradation to the multi-component enzymatic complex known as the 26S proteasome. Ubiquitin is initially produced as polyubiquitin-B or polyubiquitin-C precursor protein and its posttranslational cleavage yield multiple copies of identical 76-amino acid ubiquitin. Ubiquitin is involved in one of the most common post-translational modifications of cellular proteins, where it is linked covalently via its carboxyl terminus (Gly76) to lysine residues in target proteins. A given target lysine residue can be linked to one single ubiquitin molecule (monoubiquitylated) or to a chain of ubiquitins (polyubiquitylated). In a polyubiquitin chain, ubiquitin molecules can be linked through one of the seven lysine residues (K6, K11, K27, K29, K33, K48, and K63) or through the ubiquitin N-terminus methionine 1 residue (which generates linear chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lysine 6-linked may be involved in DNA repair; Lysine 11-linked is involved in endoplasmic reticulum-associated degradation, lysine 29-linked is involved in lysosomal degradation, and Lysine 48-linked is involved in protein degradation via the proteasome. Affimer molecules are small proteins (~ 12 kDa) that bind to target molecules with similar specificity and affinity to that of antibodies. These engineered non-antibody binding proteins are designed to mimic the molecular recognition characteristics of monoclonal antibodies in different applications. It has been shown that each affimer molecule ca bind one ubiquitin molecule and the Affimer dimerizes to bind the two ubiquitin moieties of a diubiquitin in a linkage-specific manner. The K6 Affimer is reported to tightly bind to K6 diubiquitin in a highly specific manner. It can be used to detect K6 chains and with polyubiquitin enrichment it can detect even the endogenous levels of K6 chains. (Ref.: Michel, MA., et al. (2017). Mol. Cell. 68(1); 233-246).
Specificity
Anti-diUbiquitin K6 GFP/His-tag Affimer reagent detects K6 linked poly-ubiquitin chains with a KD of approximately 20 pM. Binding to other di-ubiquitin linkages is negligible.
Immunogen
Ubiquitin Lys6 specific Affimer reagent, Anti-Ubiquitin Lys6 specific Affimer reagent, Ubiquitin Lys6 specific Affimer Reagent GFP/His tag
Application
Quality Control Testing
Evaluated by Western Blotting with Various K-linked di-ubiquitin chains.
Western Blotting Analysis (WB): A 1:1,000 dilution of this Affimer reagent detected di-ubiquitin K6-linkages.
Tested Applications
Western Blotting Analysis: A representative lot of this Affimer detected di-Ubiquitin Lys6. in Western Blotting applications (Michel, M.A., et. al. (2017). Mol Cell. 68(1):233-246).
Shouldn't it say "under license from Avacta" somewhere?
Sorry try again
https://www.merckmillipore.com/GB/en/product/Anti-Ubiquitin-Lys6-specific-Affimer-reagent-GFP-His-tag,MM_NF-MABS1918-25UG?ReferrerURL=https%3A%2F%2Fwww.bing.com%2F&bd=1
Great research Poirot.
Not sure if this is the same item
https://www.merckmillipore.com/GB/en/product/Anti-Ubiquitin-Lys6-specific-Affimer-reagent-GFP-His-tag,MM_NF-MABS1918-25UG?ReferrerURL=https%3A%2F%2Fwww.bing.com%2F&bd=1
And get this at the checkout…
1 Estimated to ship on August 09, 2021 from Merck Gillingham
https://www.sigmaaldrich.com/GB/en/sds/mm/mabs1918
Safety sheet - print date 24/7/21
Just stumbled across this site selling Affimer reagents……
https://www.sigmaaldrich.com/GB/en/product/mm/mabs1918
Sigma Aldrich are part of Merck.