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?

Name: NHE3/SLC9A3 Antibody
Brand: Novus Biologicals
SKU: NBP1-82574
Price: 549 USD

Question

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?

Accepted Answer

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.

At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Reactivity	Hu, Mu, PoSpecies Glossary
Applications	WB, ICC/IF, IHC, PAGE
Clonality	Polyclonal
Host	Rabbit
Conjugate	Unconjugated

Immunogen	This antibody was developed against Recombinant Protein corresponding to amino acids: DNPVFSPDEALDRSLLARLPPWLSPGETVVPSQRARTQIPYSPGTFCRLMPFRLSSKSVDSFLQADGPEERPPAALPEST
Isotype	IgG
Clonality	Polyclonal
Host	Rabbit
Gene	SLC9A3
Purity	Immunogen affinity purified
Innovator's Reward	Test in a species/application not listed above to receive a full credit towards a future purchase. Learn about the Innovator's Reward

Storage	Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer	PBS (pH 7.2) and 40% Glycerol
Preservative	0.02% Sodium Azide
Purity	Immunogen affinity purified

Publications using NBP1-82574	Applications	Species
Mahdi Amiri, Min Jiang, Azam Salari, Renjie Xiu, Seth L. Alper, Ursula E. Seidler Reduced surface pH and upregulated AE2 anion exchange in SLC26A3-deleted polarized intestinal epithelial cells American Journal of Physiology - Cell Physiology 2024-03-01 [PMID: 38223928]
Kim YH, Lee YK, Park SS et al. Mid-old cells are a potential target for anti-aging interventions in the elderly Nature communications 2023-11-22 [PMID: 37993434] (IHC-P, Mouse, Human)	IHC-P	Mouse, Human
Ruichao Yue, Xiaoyuan Wei, Liuyi Hao, Haibo Dong, Wei Guo, Xinguo Sun, Jiangchao Zhao, Zhanxiang Zhou, Wei Zhong Promoting intestinal antimicrobial defense and microbiome symbiosis contributes to IL-22-mediated protection against alcoholic hepatitis in mice Frontiers in Immunology 2023-10-16 [PMID: 37908362]
Mödl B, Awad M, Zwolanek D et al. Defects in microvillus crosslinking sensitize to colitis and inflammatory bowel disease EMBO reports 2023-09-11 [PMID: 37691494] (WB, ICC/IF, Mouse)	WB, ICC/IF	Mouse
Kunke, M;Knöfler, H;Dahlke, E;Zanon Rodriguez, L;Böttner, M;Larionov, A;Saudenova, M;Ohrenschall, GM;Westermann, M;Porubsky, S;Bernardes, JP;Häsler, R;Magnin, JL;Koepsell, H;Jouret, F;Theilig, F; Targeted deletion of von-Hippel-Lindau in the proximal tubule conditions the kidney against early diabetic kidney disease Cell death & disease 2023-08-26 [PMID: 37626062]
Zachos NC, Vaughan H, Sarker R et al. A NOVEL PEPTIDE PREVENTS ENTEROTOXIN- AND INFLAMMATION-INDUCED INTESTINAL FLUID SECRETION BY STIMULATING NHE3 ACTIVITY Gastroenterology 2023-07-08 [PMID: 37429363]
Peritore-Galve FC, Kaji I, Smith A et al. Increased intestinal permeability and downregulation of absorptive ion transporters Nhe3, Dra, and Sglt1 contribute to diarrhea during Clostridioides difficile infection Gut microbes 2023-06-23 [PMID: 37350393] (IHC-P, Mouse)	IHC-P	Mouse
Salari A, Zhou K, Nikolovska K et al. Human Colonoid-Myofibroblast Coculture for Study of Apical Na+/H+ Exchangers of the Lower Cryptal Neck Region International journal of molecular sciences 2023-02-21 [PMID: 36901695] (WB, ICC/IF, Human)	WB, ICC/IF	Human
Hiltz RL, McCurdy DE, Moreland S et al. Effects of weaning on regulators of volatile fatty acid absorption and intracellular pH in Holstein calves JDS Communications 2021-11-01 [PMID: 36337096] (Immunocytochemistry/ Immunofluorescence)	Immunocytochemistry/ Immunofluorescence
Donowitz M, Sarker R, Lin R et al. Identification of Intestinal NaCl Absorptive-Anion Secretory Cells: Potential Functional Significance Frontiers in Physiology 2022-07-19 [PMID: 35928564] (Block/Neutralize)	Block/Neutralize
Duclaux-Loras R, Lebreton C, Berthelet J et al. UNC45A deficiency causes microvillus inclusion disease-like phenotype by impairing myosin VB-dependent apical trafficking The Journal of clinical investigation 2022-05-16 [PMID: 35575086] (IF/IHC, ICC/IF, Human)	IF/IHC, ICC/IF	Human
Haynes J, Palaniappan B, Tsopmegha E, Sundaram U Regulation of nutrient and electrolyte absorption in human organoid-derived intestinal epithelial cell monolayers Translational Research 2022-05-01 [PMID: 35513245]
Yanda MK, Cebotaru L VX-809 mitigates disease in a mouse model of autosomal dominant polycystic kidney disease bearing the R3277C human mutation FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021-11-01 [PMID: 34662459]
Zhou K, Amiri M, Salari A Et Al. Functional characterization of the sodium/hydrogen exchanger 8 and its role in prolifeRation of colonic epithelial cells American journal of physiology. Cell physiology 2021-07-21 [PMID: 34288721]
Yue R, Wei X, Zhao J et al. Essential Role of IFN-? in Regulating Gut Antimicrobial Peptides and Microbiota to Protect Against Alcohol-Induced Bacterial Translocation and Hepatic Inflammation in Mice Frontiers in Physiology 2021-01-18 [PMID: 33536944] (Immunocytochemistry/ Immunofluorescence)	Immunocytochemistry/ Immunofluorescence
Zhang, J, Huang, Y J Et al. Primary human colonic mucosal barrier crosstalk with super oxygen-sensitive Faecalibacterium prausnitzii in continuous culture. Med (N Y) 2021-01-15 [PMID: 33511375] (IF/IHC, Human)	IF/IHC	Human
Yin J, Sunuwar L, Kasendra M et al. Fluid Shear Stress Enhances Differentiation of Jejunal Human Enteroids in Intestine-Chip Am J Physiol Gastrointest Liver Physiol 2020-10-14 [PMID: 33074011]
Klinger S Segment specific effects of resveratrol on porcine small intestinal dipeptide absorption depend on the mucosal pH and are due to different mechanisms: Potential roles of different transport proteins and protein kinases J. Nutr. Biochem. 2020-07-29 [PMID: 32738496] (WB, SDS-Page, Porcine)	WB, SDS-Page	Porcine
Hernandez-Gordillo V, Kassis T, Lampejo A et al. Fully synthetic matrices for in vitro culture of primary human intestinal enteroids and endometrial organoids Biomaterials 2020-01-01 [PMID: 32502894] (ICC/IF, Mouse, Human)	ICC/IF	Mouse, Human
Chen T, Lin R, Avula L et al. NHERF3 is Necessary for E. coli Heat Stable Enterotoxin (ST)-induced Inhibition of NHE3 - Differences in Signaling in Mouse Small Intestine and Caco-2 Cells Am. J. Physiol., Cell Physiol. 2019-07-31 [PMID: 31365292]
Pinette JA, Mao S, Millis BA et al. Brush border protocadherin CDHR2 promotes the elongation and maximized packing of microvilli in vivo Mol. Biol. Cell 2019-01-01 [PMID: 30403560] (IHC-P, Mouse)	IHC-P	Mouse
Engevik AC, Kaji I, Engevik MA et al. Loss of MYO5B Leads to Reductions in Na+ Absorption with Maintenance of CFTR-dependent Cl- Secretion in Enterocytes Gastroenterology 2018-08-22 [PMID: 30144427] (IF/IHC, Mouse)	IF/IHC	Mouse
Schlegel C, Lapierre LA, Weis VG et al. Reversible deficits in apical transporter trafficking associated with DGAT1 deficiency Traffic 2018-08-10 [PMID: 30095213] (Human)		Human
Hernandez-Gordillo V, Kassis T, Lampejo A, Choi GH Niche-inspired synthetic matrices for epithelial organoid culture bioRxiv
Zachos NC, Baetz NW, Gupta A et al. Human Rotavirus Diarrhea Is Associated with Altered Trafficking and Expression of Apical Membrane Transport Proteins bioRxiv 2019-09-26 (ICC/IF)	ICC/IF
Singh V, Yang J, Yin J et al. Cholera toxin inhibits SNX27-retromer mediated delivery of cargo proteins to the plasma membrane. J. Cell. Sci. 2018-07-20 [PMID: 30030371] (WB, Human)	WB	Human
Yin J, Tse CM, Avula LR et al. Molecular Basis and Differentiation-Associated Alterations of Anion Secretion in Human Duodenal Enteroid Monolayers Cell Mol Gastroenterol Hepatol 2018-02-09 [PMID: 29930980] (WB, Human)	WB	Human
Rajkumar P, Cha B, Yin J et al. Identifying the localization and exploring a functional role for Gprc5c in the kidney. FASEB J. 2017-12-01 [PMID: 29196502] (Mouse)		Mouse
Yin, J, Tse, C M Et al. A common NHE3 single-nucleotide polymorphism has normal function and sensitivity to regulatory ligands. Am J Physiol Gastrointest Liver Physiol 2017-08-01 [PMID: 28495802] (FLOW, Human)	FLOW	Human
Foulke-Abel J, In J, Yin J et al. Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology. Gastroenterology. 2015-12-08 [PMID: 26677983] (WB, IHC-Fr, Human)	WB, IHC-Fr	Human
Engevik MA, Engevik KA, Yacyshyn MB et al. Human Clostridium difficile infection: Inhibition of NHE3 and microbiota profile. Am. J. Physiol. Gastrointest. Liver Physiol. 2014-12-31 [PMID: 25552580]
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