Simple Western: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Simple Western lane view shows a specific band for NHE3 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions ...read more
Hu, Mu, Rt, Po, Ma-Op, Pm, RbSpecies Glossary
WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IP, KD, KO
This NHE3 [p Ser552] (14D5) antibody is useful in ELISA, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunoprecipitation and Western Blot, where a band can be seen at approx. 85 kDa.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. Use in FLOW reported in scientific literature (PMID: 21255452). Knockdown and knockout Validated reported in scientific literature (PMID: 31276916).
85 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Read 1 Review rated 4 using NB110-81529 in the following applications:
Porcine reactivity reported in scientific literature (PMID: 29510506).
Packaging, Storage & Formulations
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Tris-Glycine and 0.15M NaCl
0.05% Sodium Azide
Protein G purified
Alternate Names for NHE3/SLC9A3 [p Ser552] Antibody (14D5)
solute carrier family 9 (sodium/hydrogen exchanger), member 3
Solute carrier family 9 member 3
Na+/H+ exchanger (NHE) family members are integral membrane proteins which play a key role in catalyzing the extrusion of intracellular proton (H+) ions in exchange for extracellular sodium (Na+) ions for regulation of cellular pH as well as transepithelial ion/water transport. NHE3 is one of the best characterized NHEs that is a major proton extruding system driven by inward Na+ ion chemical gradient and is implicated in pH regulation to eliminate acids generated during metabolism or to tackle any potential adverse conditions in tissue microenvironment. NHE3 exists as a multi-pass membrane protein in cell membranes and is expressed most abundantly in colon and the small intestine, followed by the kidneys and the stomach, as well as in ventrolateral region of the medulla oblongata. In intestinal epithelial cells, it localizes to the ileal brush border and SGK1 mediated phosphorylation at Ser-661 has been shown to be associated with its enhanced cell membrane localization. NHE3 binds to SLC9A3R1 and SLC9A3R2, and interacts with CHP1, CHP2, SHANK2 PDZD3 and (interactions may decrease in response to elevated calcium ion levels). NHE3 is regulated by several hormones, neurotransmitters, and associated signaling systems such as cAMP, cGMP, and elevated intracellular calcium via mechanisms that are largely partially understood. NHE3 knock-out mice suffers from chronic diarrhea and altered salt as well as water homeostasis.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for NHE3/SLC9A3 Antibody (NB110-81529). (Showing 1 - 2 of 2 FAQs).
What application was used to test on mouse species? Was it WB, ELISA, IHC-paraffin, IHC-frozen, ICC/IF, or etc?
We tested each lot in Western blot on human, mouse, and rat colon, small intestine and kidney samples. There were bands expressed around 25kDa and 50kDa, with weaker bands at approx. 80-90 kDa observed. This antibody is guaranteed to work in samples from mouse that are expected to express the protein. Since it is a phospho specific antibody the customer must use BSA to block and not milk.
The molecular weight of NHE3 is 92kDa but I see a band above 100 kDa. I am confused, because the size of these protein should not be greater than 100KD. Can you please help explain?
Higher than predicted molecular weight seen on WB is not uncommon if the protein is posttranslationally modified, especially glycosylated, and such is the case for NHE3. Please note that NHE3 gets phosphorylated and glycosylated (Uniprot ID P48764) and potentially, these modifications are adding ~10kDa extra to the predicted molecular weight. If you want to confirm it 100%, you could try de-glycosylating your protein and then performing a WB on it.
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