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
Product Details
Summary
Reactivity
Hu, Mu, Rt, Po, Ma-Op, Pm, RbSpecies Glossary
Applications
WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IP, KD, KO
Knockdown Validated reported in scientific literature (PMID 31276916)
Knockout Validated reported in scientific literature (PMID 31276916)
Simple Western 1:1000
Western Blot 1-5 ug/ml
Application Notes
In Western Blot, 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.
Theoretical MW
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.
Reviewed Applications
Read 1 Review rated 4 using NB110-81529 in the following applications:
Porcine reactivity reported in scientific literature (PMID: 29510506).
Packaging, Storage & Formulations
Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tris-Glycine and 0.15M NaCl
Preservative
0.05% Sodium Azide
Concentration
2 mg/ml
Purity
Protein G purified
Alternate Names for NHE3/SLC9A3 [p Ser552] Antibody (14D5)
isoform 3
MGC126718
NHE3
NHE3MGC126720
SLC9A3
solute carrier family 9 (sodium/hydrogen exchanger), member 3
Solute carrier family 9 member 3
Background
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.
Limitations
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.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
![Western Blot: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Detection of NHE3 [p Ser552] in human kidney lysate.](http://images.novusbio.com/fullsize/NHE3-SLC9A3-p-Ser552-Antibody-14D5-Western-Blot-NB110-81529-img0005.jpg "Western Blot: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Detection of NHE3 [p Ser552] in human kidney lysate.")
![Immunohistochemistry PTH Antibody (918462) [Unconjugated]](https://images.novusbio.com/images2/PTH_MAB7665_Immunohistochemistry_16650.jpg)
![Western Blot PTH Antibody (918462) [Unconjugated]](https://images.novusbio.com/images2/PTH_MAB7665_Western_Blot_16656.jpg)
![NHE3/SLC9A3 [p Ser552] Antibody (14D5)](/sites/all/modules/enterprise-tech/et_datasheets/images/novus_guarantee.png "NHE3/SLC9A3 [p Ser552] Antibody (14D5)")
![Immunohistochemistry: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Mouse intestine, image courtesy of verified customer review.](http://images.novusbio.com/fullsize/NHE3-SLC9A3-p-Ser552-Antibody-14D5-Immunofluorescence-NB110-81529-img0008.jpg "Immunohistochemistry: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Mouse intestine, image courtesy of verified customer review.")
![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 using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/NHE3-SLC9A3-p-Ser552-Antibody-14D5-Simple-Western-NB110-81529-img0006.jpg "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 using the 12-230 kDa separation system.")
![Immunohistochemistry CFTR Antibody (24-1) [Unconjugated] - C-terminus](https://images.novusbio.com/images2/CFTR_MAB25031_Immunohistochemistry_6973.jpg)
![Immunohistochemistry CFTR Antibody (24-1) [Unconjugated] - C-terminus](https://images.novusbio.com/images2/CFTR_MAB25031_Immunohistochemistry_9671.jpg)
![Immunoprecipitation CFTR Antibody (24-1) [Unconjugated] - C-terminus](https://images.novusbio.com/images/mab25031_human-cftr-c-terminus-mab-clone-24-1-41202412513815.jpg)
![Simple Western DPPIV/CD26 Antibody [Unconjugated]](https://images.novusbio.com/images2/DPPIV_AF1180_Simple_Western_20720.jpg)
![Immunohistochemistry DPPIV/CD26 Antibody [Unconjugated]](https://images.novusbio.com/images2/DPPIV_AF1180_Immunohistochemistry_17209.jpg)
![Immunocytochemistry DPPIV/CD26 Antibody [Unconjugated]](https://images.novusbio.com/images2/DPPIV_AF1180_Immunocytochemistry__Immunofluorescence_17210.jpg)
