Reagenzien und Instrumente für die Immunologie Zellbiologie und Molekularbiologie

Anti-NCC (Thiazide sensitive NaCl cotransporter) (Thr53) Antibody

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Katalog-Nummer p1311-53

Size : 100uL

Marke : Phosphosolutions

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Anti-NCC (Thiazide sensitive NaCl cotransporter) (Thr53) Antibody

Our Anti-NCC (Thiazide sensitive NaCl cotransporter) (Thr53) rabbit polyclonal phosphospecific primary antibody from PhosphoSolutions is produced in-house. It detects human, mouse, and rat NCC (Thiazide sensitive NaCl cotransporter) and is antigen affinity purified from pooled serum. It is great for use in WB, IHC.

Western blot of mouse kidney lysate showing specific immunolabeling of the ~160 kDa NCC protein phosphorylated at Thr53 in the first lane (-). Phosphospecificity is shown in the second lane (+) where immunolabeling is completely eliminated by blot treatment with lambda phosphatase (λ-Ptase, 1200 units for 30 min).

Immunostaining of PFA perfused frozen kidney sections from WT and NCC KO mice showing specific labeling of the NCC protein phosphorylated at Thr53 (Cat. p1311-53, red, 1:100,000) on the left and the absence of staining in the KO on the right. (Image courtesy of Lauren Miller, Ellison Lab, OHSU.)

Two Western blots show the abundance of pNCCT53 (Cat. p1311-53, 1:200) and tNCC in male Kcnj10fl/fl mice and in Ks-Kir4.1–KO mice treated with vehicle (control) and FK506 (0.75 mg/kg BW). Image from publication CC-BY-4.0. PMID: 36821372

Immunostaining of mouse kidney sections from control (CTL) and BRL37344 treated (BRL, 1 mg/kg) specifically labeling localized phosphorylated NCC thr53 (Cat. p1311-53, green) and counterstained with Evans Blue (in red). Image from publication. CC-BY-4.0, PMID: 38652212

Immunolabeling of mouse kidney paraffin section specifically detecting NCC Thr53 (Cat. p1311-53, red, 1:1000) phosphorylation with the increased NCC activation in Ang-II-infused Prdx5 KO kidney. Image from publication, CC-BY-4.0. PMID: 39857434.

Western blot image of kidney measuring levels of pNCC-T53 and total NCC in Kir4.2+/+ and Kir4.2−/− mice. Image from publication, CC-BY-4.0. PMID: 38886379.

Western blot of mouse kidney lysate showing specific immunolabeling of the ~160 kDa NCC protein phosphorylated at Thr53 in the first lane (-). Phosphospecificity is shown in the second lane (+) where immunolabeling is completely eliminated by blot treatment with lambda phosphatase (λ-Ptase, 1200 units for 30 min).

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Species Reactivity

Mouse

Applications

IHC, WB

Host Species

Rabbit

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Data Sheet

KO Validated

Trial Size

Pooled Serum

SKU: p1311-53

Ships: 1-2 business days

Product Details

Target

NCC (Thiazide sensitive NaCl cotransporter)

Scientific Background

The thiazide-sensitive sodium chloride cotransporter, NCC, is the major NaCl transport protein in the distal convoluted tubule (DCT) and plays an important role in maintaining blood pressure (Rosenbaek et al., 2014, Feng et al., 2015). Phosphorylation of NCC at Thr-53, Thr-58, and Ser-71 is an essential mediator of NCC function (Rosenbaek et al., 2014). NCC is constitutively cycled to the plasma membrane, and upon stimulation, it can be phosphorylated to both increase NCC activity and decrease NCC endocytosis, together increasing NaCl transport in the DCT (Feng et al., 2015).

Format

Antigen Affinity Purified from Pooled Serum

Clonality

Polyclonal

Isotype

IgG

Applications

IHC, WB

Host Species

Rabbit

Gene Name

SLC12A3

Molecular Weight

160 kDa

Immunogen

Synthetic phospho-peptide corresponding to amino acid residues surrounding Thr53 of mouse NCC, conjugated to keyhole limpet hemocyanin (KLH).

Immunogen Species

Mouse

Species Reactivity

Mouse

Expected Reactivity

Guinea Pig, Hamster

Antibody Registry ID

AB_2650477

Storage

Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.

Physical State

Liquid

Production Notes

Prepared from pooled rabbit serum by affinity purification via sequential chromatography on phospho and non-phosphopeptide affinity columns.

Buffer

10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg per ml BSA and 50% glycerol.

Dilution Ranges

WB: 1:1000-1:6000
IHC: 1:100-1:10,000

Conjugate

Unconjugated

Specificity

Specific for endogenous levels of the ~160 kDa NCC protein phosphorylated at Thr53. Band of interest smearing likely due to glycosylation. Immunolabeling is completely eliminated by treatment with l-phosphatase.

Post Translational Modification Type

Phosphorylated

Post Translational Modification Residue

Thr53

Quality Control

Western blots performed on each lot.

Usage Statement

For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.

Expiration

After date of receipt, stable for at least 1 year at -20°C.

Synonyms

FLJ96318 antibody, Na Cl cotransporter antibody, Na Cl symporter antibody, Na-Cl symporter antibody, NaCl electroneutral thiazide sensitive cotransporter antibody, NCCT antibody, SLC12A3 antibody, S12A3_HUMAN antibody, slc12a3 antibody, Solute carrier family 12 (sodium/chloride transporters) member 3 antibody, Solute carrier family 12 member 3 antibody, Thiazide sensitive Na Cl cotransporter antibody, Thiazide sensitive sodium chloride cotransporter antibody, Thiazide-sensitive sodium-chloride cotransporter antibody, TSC antibody

UniProt ID

P59158

NCBI Gene ID

20497

Shipping

Blue Ice

Western Blot Protocol: Download

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