Anti-SOCS3 抗体 (ab723)

Thank you for your email. Ab723 is 325 Euros and is in stock in our office in Great Britain so upon ordering, you should receive it within a couple of days. You can place your order online, or via fax, email or phone. Below is the appropriate contact details: Abcam Ltd 21 Cambridge Science Park Milton Road Cambridge CB4 0TP UK Email: orders@abcam.com Phone: +44 1223 472030 Fax: +44 1223 472038 Office hours: 08.30 - 17.00 hrs (UK) Monday to Friday Please let us know how ab723 works out for you and contact me again if you have any more questions.

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soon as any new information is brought to our attention. As far as we are aware, cross reactivity with rat has not yet been tested for use with ab723. The antibody cross-reacts with human and is expected to cross-react with rat and mouse due to sequence homology. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. Please let me know if you require any further assistance.

Thank you for the details that you have provided. We have had no other complaints concerning this antibody. The antibody's originator has provided me with the following protocol that they used when testing the antibody which should be helpful for you: - Lysis. Cell pellets were washed with ice-cold PBS. 1 ml of RIPA buffer was added per 1E8 cells and incubated on ice for 20 min, vortexing 2-3 times, briefly. The lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with BioRad protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was then added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton -100, 1% Sodium deoxycholate, 0.1% SDS). - SDS PAGE. Samples were run at 200V constant on a 12% acrylamide SDS-PAGE mini gel - using Biorad Mini-Protean 3 kit and protocols. Before loading samples had 5% (v/v) 2-ME added and were boiled for 3 minutes. - Transfer. We used a Biorad Mini Trans-Blot, constant 100 V for 1 hour. Transfer Buffer was 20 mM Tris pH 8.0, 150 mM Glycine, 10% Methanol. We transferred to Millipore PVDF membrane and stained with Ponceau Red to evaluate the transfer. - Staining. The membrane was blocked in 2.5% skimmed milk in TBS-T (TBS + 0.05% Tween-20) for 1 hr at room temperature with agitation. Primary antibody was incubated for 1 hr at room temperature with agitation. We used sigma secondary (Sigma anti-goat-HRP Product # A4174, use 1:3000) for 1 hr at room temperature with agitation. We washed with TBST three times after primary and secondary antibody, each wash lasting for 5-10 mins. ECL-plus (Amersham) was used rather than ECL, which is considerably more sensitive. Final detection was on autoradiography film. As you got no signal with the positive control, MOLT-4 lysate, I am willing to give you a replacement batch of this antibody if you would like. Please let me know what you would like to do.