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Product Name
Anti-TMEM163 antibody
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Description
Rabbit polyclonal antibody to TMEM163
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Tested applications
WB, ICC, IHC-P, FC
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Species reactivity
Human, Mouse, Rat
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Alternative names
DC29 antibody; SV31 antibody
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Isotype
Rabbit IgG
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Preparation
This antigen of this antibody was synthetic peptide within human tmem163 aa 50-100.
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Clonality
Polyclonal
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Formulation
Liquid, 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
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Storage instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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Applications
WB:1:500-1:2,000
ICC:1:50-1:200
IHC-P:1:50-1:200
FC:1:50-1:100
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Validations
Fig1:; 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 ( 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: Human mouse colon tissue lysate; Predicted band size: 31 kDa; Observed band size: 38 kDa
Fig2:; 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 ( 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).
Fig3:; 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 ddH; 2; O and PBS, and then probed with the primary antibody ( 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.
Fig4:; 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 ddH; 2; O and PBS, and then probed with the primary antibody ( 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.
Fig5:; Flow cytometric analysis of TMEM163 was done on F9 cells. The cells were fixed, permeabilized and stained with the primary antibody ( 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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References
- 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.
- Sanchez VB. et. al. Transmembrane 163 (TMEM163) protein effluxes zinc. Arch Biochem Biophys. 2019 Nov 30;677:108166.
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