The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 40, 55 kDa (predicted molecular weight: 55 kDa).
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This is one of the several different receptors for 5-hydroxytryptamine (serotonin), a biogenic hormone that functions as a neurotransmitter, a hormone, and a mitogen. This receptor is a ligand-gated ion channel, which when activated causes fast, depolarizing responses in neurons. It is a cation-specific, but otherwise relatively nonselective, ion channel.
Expressed in cerebral cortex, amygdala, hippocampus, and testis. Detected in monocytes of the spleen and tonsil, in small and large intestine, uterus, prostate, ovary and placenta.
Belongs to the ligand-gated ion channel (TC 1.A.9) family. 5-hydroxytryptamine receptor (TC 1.A.9.2) subfamily. HTR3A sub-subfamily.
Western blot - Anti-5HT3A receptor antibody (ab45314)
All lanes : Anti-5HT3A receptor antibody (ab45314) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466) Lane 2 : Human testis tissue lysate - total protein (ab30257) Lane 3 : Human heart tissue lysate - total protein (ab29431) Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
In heart and skeletal muscle, ab45314 detects a band at 55 kDa, corresponding to the predicted molecular weight of 5HT3A. We are unsure as to why a lower band of 40 kDa is seen in brain and testis. However, subsequent re-testing of this antibody on testis produced a single band at 55 kDa, suggesting that the smaller band originally observed may be due to lysate variation.
Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
IHC image of 5HT3A staining in human hippocampus FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45314, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Ozcan CU et al. Evaluation of the relation between interstitial cells of cajal (CD117) and serotonin receptor (5HT-3A) with postfundoplication dysphagia. Int J Surg13:137-41 (2015).
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