Read MoreĪlors pour la séquence de SERT chez la moule que j’étudie, elle n’est pas connu (Elliptio Complanata).
SEROTONIN TRANSPORTER CODE
1) Abcam product code ab102048 2) Abcam order reference number or product batch number, 1148703 3) Description of the problem: no bands was detected at all 4) Sample preparation: Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysate Lysis buffer, RIPA Protease inhibitors: Fisher protease and phosphatase inhibitor Phosphatase inhibitors: Reducing agent, DTT and beta-ME Boiling for ≥5 min? yes Protein loaded ug/lane or cells/lane, 20 ug Positive control, self made serotonin transporter (SERT) antibody Negative control, HEK cell lysate without SERT overexpression 5) Percentage of gel: 8% Type of membrane: nitrocellulose Protein transfer verified, yes Blocking agent and concentration: 5% milk Blocking time: 30 min Blocking temperature: RT 6) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution: 1: 300-500 Diluent buffer: 5% BSA Incubation time: Overnight Incubation temperature: 4 degree 7) Secondary antibody: Species: Goat Reacts against: rabbit Concentration or dilution: 1: 1000 Diluent buffer: 5% milk Incubation time: 1h Incubation temperature: RT Fluorochrome or enzyme conjugate: HRP 8) Washing after primary and secondary antibodies: 3 times 5 min Buffer: TBS+Tween (0.0005%) Number of washes: 3 9)Detection method 10) How many times have you run this staining? 3 times Do you obtain the same results every time? Yes, no signal at all What steps have you altered to try and optimize the use of this antibody? Increasing antibody concentration, or block with BSA instead of milk Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem. Wish I could hear from you soon and have your suggestions, thank you very much. Please find attached the questionaire as well as the images you wanted in ppt, thanks.