NHE3/SLC9A3 [p Ser552] Antibody (14D5)

Images

 
Western Blot: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Detection of NHE3 [p Ser552] in human kidney lysate.
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
Immunofluorescence: NHE3/SLC9A3 [p Ser552] Antibody (14D5) [NB110-81529] - Mouse intestine, image courtesy of verified customer review.

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Op, Pm, RbSpecies Glossary
Applications WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IP, ICC/IF, KD, KO
Clone
14D5
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
2 mg/ml

NHE3/SLC9A3 [p Ser552] Antibody (14D5) Summary

Immunogen
Synthetic peptide made to a region surrounding the Serine 552 portion of rat NHE3. [UniProt# P26433]
Modification
p Ser552
Specificity
This antibody detects NHE3 posphorylated at Serine 552, and does not cross-react with the non-phosphopeptide.
Isotype
IgG2a
Clonality
Monoclonal
Host
Mouse
Gene
SLC9A3
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1-5 ug/ml
  • Simple Western 1:1000
  • ELISA 1:100-1:2000
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:50-1:1000
  • Immunohistochemistry 1:50-1:1000
  • Immunoprecipitation 1:10-1:500
  • Immunofluorescence
  • Knockdown Validated
  • Knockout Validated
Application Notes
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).
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:

Publications
Read Publications using
NB110-81529 in the following applications:

Reactivity Notes

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.

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Publications for NHE3/SLC9A3 Antibody (NB110-81529)(13)

We have publications tested in 6 confirmed species: Human, Mouse, Rat, Opossum, Porcine, Primate.

We have publications tested in 8 applications: ELISA, FLOW, ICC/IF, IHC, IP, Knockdown Validated, Knockout Validated, WB.


Filter By Application
ELISA
(1)
FLOW
(1)
ICC/IF
(3)
IHC
(1)
IP
(4)
Knockdown Validated
(1)
Knockout Validated
(1)
WB
(11)
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Human
(4)
Mouse
(3)
Rat
(4)
Opossum
(2)
Porcine
(2)
Primate
(2)
All Species
Showing Publications 1 - 10 of 13. Show All 13 Publications.
Publications using NB110-81529 Applications Species
Koizumi M, Ueda K, Niimura F et al. Podocyte Injury Augments Intrarenal Angiotensin II Generation and Sodium Retention in a Megalin-Dependent Manner Hypertension Sep 1 2019 [PMID: 31352823]
Hayasaki T, Ishimoto T, Doke T et al. Fructose increases the activity of sodium hydrogen exchanger in renal proximal tubules that is dependent on ketohexokinase J. Nutr. Biochem. Jun 8 2019 [PMID: 31276916] (WB, Knockdown Validated, Knockout Validated, Porcine, Mouse) WB, Knockdown Validated, Knockout Validated Porcine, Mouse
Masuda T, Watanabe Y, Fukuda K et al. Unmasking a sustained negative effect of SGLT2 inhibition on body fluid volume in the rat Am. J. Physiol. Renal Physiol. May 23 2018 [PMID: 29790389] (Rat) Rat
Klinger S, Breves G. Resveratrol Inhibits Porcine Intestinal Glucose and Alanine Transport: Potential Roles of Na+/K+-ATPase Activity, Protein Kinase A, AMP-Activated Protein Kinase and the Association of Selected Nutrient Transport Proteins with Detergent Resistant Membranes Nutrients. Mar 3 2018 [PMID: 29510506] (WB, Porcine) WB Porcine
Fenton RA, Poulsen SB, de la Mora Chavez S et al. Renal tubular NHE3 is required in the maintenance of water and sodium chloride homeostasis. kidney Int. Aug 1 2017 [PMID: 28385297] (WB, Mouse) WB Mouse
Ozkucur N, Song B, Bola S et al. NHE3 phosphorylation via PkCEta marks the polarity and orientation of directionally migrating cells. Cell. Mol. Life Sci. 2014 May 01 [PMID: 24788043] (WB, IP, ICC/IF, Human) WB, IP, ICC/IF Human
Perike S, Ozkucur N, Sharma P et al. Phospho-NHE3 forms membrane patches and interacts with beta-actin to sense and maintain constant direction during cell migration. Exp. Cell Res. 3/19/2014 [PMID: 24657527] (IP, WB, Human) IP, WB Human
Liu T, Jose PA. Gastrin Induces Sodium-Hydrogen Exchanger 3 Phosphorylation and mTOR Activation via a Phosphoinositide 3-Kinase-/Protein Kinase C-Dependent but AKT-Independent Pathway in Renal Proximal Tubule Cells Derived From a Normotensive Male Human. Endocrinology 2013 Feb [PMID: 23275470] (WB, Human) WB Human
Rieg T, Gerasimova M, Murray F et al. Natriuretic effect by exendin-4, but not the DPP-4 inhibitor alogliptin, is mediated via the GLP-1 receptor and preserved in obese type 2 diabetic mice Am J Physiol Renal Physiol 2012 Oct [PMID: 22832924] (WB, Mouse) WB Mouse
Ozkucur N, Perike S, Sharma P et al. Persistent directional cell migration requires ion transport proteins as direction sensors and membrane potential differences in order to maintain directedness. BMC Cell Biol. 2011 Jan [PMID: 21255452] (FLOW, ICC/IF, IP, WB, Rat, Human) FLOW, ICC/IF, IP, WB Rat, Human
Show All 13 Publications.

Review for NHE3/SLC9A3 Antibody (NB110-81529) (1) 41

Average Rating: 4
(Based on 1 review)
We have 1 review tested in 1 species: Mouse.

Reviews using NB110-81529:
Filter by Applications
IHC-P
(1)
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Mouse
(1)
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Images Ratings Applications Species Date Details
Immunohistochemistry-Paraffin NHE3/SLC9A3 NB110-81529
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reviewed by:
Anjum Jafri
IHC-P Mouse 03/16/2017
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedMouse Intestine
SpeciesMouse

Product General Protocols

View specific protocols for NHE3/SLC9A3 Antibody (NB110-81529): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for NHE3/SLC9A3 Antibody (NB110-81529). (Showing 1 - 2 of 2 FAQs).

  1. 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.
  2. 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. 

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB110-81529AF350
Alexa Fluor 405 NB110-81529AF405
Alexa Fluor 488 NB110-81529AF488
Alexa Fluor 532 NB110-81529AF532
Alexa Fluor 594 NB110-81529AF594
Alexa Fluor 647 NB110-81529AF647
Alexa Fluor 700 NB110-81529AF700
Alexa Fluor 750 NB110-81529AF750
Biotin NB110-81529B
DyLight 350 NB110-81529UV
DyLight 405 NB110-81529V
DyLight 488 NB110-81529G
DyLight 550 NB110-81529R
DyLight 594 NB110-81529DL594
DyLight 650 NB110-81529C
DyLight 680 NB110-81529FR
DyLight 755 NB110-81529IR
FITC NB110-81529F
HRP NB110-81529H
Janelia Fluor 549 NB110-81529JF549
Janelia Fluor 646 NB110-81529JF646

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Recent Reviews

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Anjum Jafri
03/16/2017
Application: IHC-P
Species: Mouse

Bioinformatics

Gene Symbol SLC9A3