Synthetic peptide within Human TMEM163 aa 55-104 / 289.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Target Molecular Weight
Predicted band size 31 kDa
Positive Control
Human skin tissue lysate, human small intestine tissue lysate, human mouse colon tissue lysate, 293T, rat large intestine tissue, human prostate carcinoma tissue, F9.
Previously, it was shown that purified rodent Tmem163 protein reconstituted in liposomes forms a dimer and transports zinc in a proton-dependent manner. It was also shown that Tmem163's ability to bind and transport zinc becomes inactivated upon alanine substitution of two species-conserved aspartate residues at positions 123 and 127. Thus, the data indicate that these double aspartate residues are zinc binding sites, but that they do not affect Tmem163 dimerization. Note, however, that this particular study did not address the question of whether the protein serves to transport zinc in or out of living cells. TMEM163 may play a crucial role in cellular zinc homeostasis by extruding cytoplasmic zinc ions to the extracellular environment. Noteworthy is that TMEM163 is unique in that it has a short C-terminus region, whereas the human ZNT family of efflux transporters have typically long C-terminus regions. Despite such structural difference, data from both functional and phylogenetic studies of TMEM163 suggest that it should now be referred to as ZNT11 protein and its gene named as SLC30A11.
Background References
1. Chakraborty S. et. al. Role of Tmem163 in zinc-regulated insulin storage of MIN6 cells: Functional exploration of an Indian type 2 diabetes GWAS associated gene. Biochem Biophys Res Commun. 2020 Feb 19;522(4):1022-1029.
2. Sanchez VB. et. al. Transmembrane 163 (TMEM163) protein effluxes zinc. Arch Biochem Biophys. 2019 Nov 30;677:108166.
Sequence Similarity
Belongs to the TMEM163 family.
Subcellular Location
Early endosome membrane, synaptic vesicle membrane.
Western blot analysis of TMEM163 on different lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1901-91, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Human skin tissue lysate Lane 2: Human small intestine tissue lysate Lane 3: Mouse colon tissue lysate
Predicted band size: 31 kDa Observed band size: 38 kDa
ICC staining of TMEM163 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded rat large intestine tissue using anti-TMEM163 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-91, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-TMEM163 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-91, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of TMEM163 was done on F9 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-91, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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