The kidneys play a major role in the regulation of glucose levels. Kidneys filter approx. 180 g of glucose per day from the blood, and this is mostly reabsorbed back into the blood in the proximal tubules. Typically, glucose is first absorbed within epithelium by a specific transporter protein, Sodium glucoseco transporters (SGLT), in the brush-border membrane and then it is transported out of the cell across the basolateral membranes by a facilitated sugar transporter (GLUTs). At least 3 members of SGLTs (SGLT1-3) have been cloned and characterized from various species. Individual member of this family have identical predicted secondary structures with up to 14 transmembrane domains. SGLT1-3 genes code for protein of approx 659-672 residues (calculated size of ~75kD). Both N and C-termini are predicted to be extracellualr. There is approx 60-70% homology between SGLT1-3. SGLTs transport a-methyl-D-glucoside (a-MDG), a non-metabolized model substrate, in Na-dependent manner. SGLT1 does not discriminate a-MDG, glucose, and galactose. SGLT2/3 do not transport D-galactose efficiently.
SGLT1/NAGT or SLC5A1/NAGT (rat/mouse 665 aa; human 664 aa, chromosome 22q13.1, ~75kD) is a high affinity, Na+-coupled, intestinal responsible for active glucose transport across the brush border membrane. In the kidney, SGLT1 is expressed in proximal tubule Sq1 segments. It is also expressed in the intestine. Defects in SGLT1 gene have been implicated in congenital glucose-galactose malabsorption syndrome (GGM). SGLT2/SLC5A2 (rat/mouse 670 aa; human 672 aa, chromosome 16p11.2) is the low affinity, high capacity Na+-glucose transporter located in the S1 segments of proximal tubules. It is ~60% identical with SGLT1. SGLT2 mediates saturable Na-dependent and phlorizin-sensitive glucose transport. In contrast with SGLT1, SGLT2 does not transport D-galactose. Defect in SGLT2 may be associated with renal glycosuria. SGLT3/SLC54 (pig 660 aa; mouse 656/660/616 aa; human 659 aa, chromosome 22), originally named SAAT1 or pSGLT2, was initially identified in LLC-Pk1 cell line derived from pig renal epithelium. It is also low affinity Na-glucose transporter. It is expressed in kidney, intestine, liver, skeletal muscle and spleen. Like SGLT2, SGLT3 has a low affinity for sugars, and is highly selective for D-glucose and low affinity for D-galactose.
Applications
Suitable for use in ELISA, Western Blotting. Other applications not tested.
Recommended Dilution
Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure antibody using ECL.
ELISA: 1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity pure. Control peptide can be used to coat ELISA plates at 1ug/ml. Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
A 14 AA peptide sequence from the C-terminal, cytoplasmic domain 6 of pig SGLT-3 (1) was synthesized, coupled to KLH
Form
Supplied as a liquid, neat serum, 40% glycerol.
Specificity
The porcine peptide sequence is poorly conserved in other species SGLT3. No significant sequence homology exists with other SGLTs. For mouse SGLT3, we recommend the use antibody and SG-32 for human SGLT-3 that are made to the mouse and human SGLT-3 peptide, respectively. Control peptide, because of its low MW (<3kD), is not suitable for Western. It should be used for ELISA or antibody blocking experiments (use 5-10 ug control peptide per 1 ug of aff pure IgG or 1 ul antiserum) to confirm antibody specificity . This antibody has previsouly been available for porcine SGLT-2. The same antibody is now reclassified as porcine SGLT-3.
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