This antibody gave a positive signal in the following rat tissue lysates:
Cortex; Hippocampus; Spinal Cord.
This antibody gave a positive result in IHC in the following FFPE tissue: Rat normal pancreas.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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 52 kDa (predicted molecular weight: 52 kDa).
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use at an assay dependent concentration.
Functions as a sodium-dependent amino acid transporter which countertransport protons. Mediates the saturable, pH-sensitive, and electrogenic cotransport of several neutral amino acids including glycine, asparagine, alanine, serine, glutamine and histidine with sodium.
Predominantly expressed in stomach, brain, liver, lung and intestinal tract.
Belongs to the amino acid/polyamine transporter 2 family.
IHC image of SLC38A5 / SNAT5 staining in Rat normal pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab72717, 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-SLC38A5 / SNAT5 antibody (ab72717)
All lanes : Anti-SLC38A5 / SNAT5 antibody (ab72717) at 1 µg/ml
Lane 1 : Cortex (Rat) Tissue Lysate Lane 2 : Hippocampus (Rat) Tissue Lysate Lane 3 : Spinal Cord (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 52 kDa Observed band size: 52 kDa Additional bands at: 28 kDa, 30 kDa, 40 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab72717 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72717, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Day PE et al. Partitioning of glutamine synthesised by the isolated perfused human placenta between the maternal and fetal circulations. Placenta34:1223-31 (2013).
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