Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)

Rabbit Recombinant Monoclonal SLC12A1/NKCC2 antibody. Suitable for WB, IHC-P and reacts with Human, Mouse, Rat, Recombinant fragment - Mouse samples.

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Images

Multiplex immunohistochemistry - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (AB313640), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)	IP
Human	Tested	Tested	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Not recommended	Not recommended	Not recommended
Rat	Tested	Tested	Not recommended	Not recommended	Not recommended
Recombinant fragment - Mouse	Tested	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Recombinant fragment - Mouse	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat, Recombinant fragment - Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat, Recombinant fragment - Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat, Recombinant fragment - Mouse	Dilution info -	Notes -

Associated Products

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1 product for Alternative Version

Target data

See full target information Slc12a1

Function

Renal sodium, potassium and chloride ion cotransporter that mediates the transepithelial NaCl reabsorption in the thick ascending limb and plays an essential role in the urinary concentration and volume regulation. Electrically silent transporter system. Isoform A. High affinity, high capacity cotransporter for sodium, potassium and chloride ions, with a coupling ratio 1Na(+):1K(+):2Cl(-). Isoform B. High affinity, low capacity cotransporter for sodium, potassium and chloride ions, with a coupling ratio 1Na(+):1K(+):2Cl(-). Isoform F. Low affinity, low capacity cotransporter for sodium, potassium and chloride ions, with a coupling ratio 1Na(+):1K(+):2Cl(-).

Alternative names

Nkcc2, Slc12a1, Solute carrier family 12 member 1, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1, Kidney-specific Na-K-Cl symporter, Na-K-2Cl cotransporter 2, BSC1, mNKCC2

Recommended products

Rabbit Recombinant Monoclonal SLC12A1/NKCC2 antibody. Suitable for WB, IHC-P and reacts with Human, Mouse, Rat, Recombinant fragment - Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR28176-90
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The SLC12A1 protein also known as the NKCC2 transporter is an important player in ion transport within renal physiology. With a molecular weight of around 120 kDa NKCC2 functions in the reabsorption of sodium potassium and chloride ions a function facilitated by its location in the thick ascending limb of the loop of Henle in the kidneys. This location and functionality are critical for renal concentration mechanisms and contribute to the maintenance of electrolyte and fluid balance.

Biological function summary

This transporter plays an essential role in concentrating urine and regulating blood pressure. NKCC2 is a part of the larger solute carrier family and does not usually form complexes with other proteins. It facilitates the trans-epithelial movement of Na+ K+ and 2Cl- ions from the lumen into renal epithelial cells which is fundamental for the creation of an osmotic gradient essential for water reabsorption.

Pathways

The SLC12A1 protein is a significant component of renal electrolyte transport and blood pressure regulation pathways. It is intricately involved in the renin-angiotensin-aldosterone system (RAAS) which regulates blood volume and systemic vascular resistance. In these pathways NKCC2's activity tightly connects it to other ion channel proteins and transporters such as ENaC which also play roles in blood pressure regulation by influencing sodium handling in the nephron.

Associated diseases and disorders

Dysregulation of NKCC2 function can result in Bartter syndrome and hypertension. Bartter syndrome a rare disorder characterized by salt wasting and electrolyte imbalances stems from mutations affecting NKCC2. This transporter also impacts hypertension where its dysregulation can alter sodium reabsorption processes linking it to other proteins involved in blood pressure control such as the mineralocorticoid receptor. Understanding the role of NKCC2 provides insight into potential therapeutic targets for these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

Multiplex immunohistochemistry - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining GLEPP1/PTPRO with Anti-GLEPP1/PTPRO antibody [EPR28645-553] ab322047 at a 1:2000 (0.261 ug/ml) dilution, ab313640 anti-SLC12A1 used at 1:2000 (0.253 ug/ml) dilution and Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1:4000 (0.141 ug/ml) dilution.
Panel A: merged staining of anti-PTPRO (magenta; Opal™690), anti-SCL12A1 (green; Opal™520) and anti-Angiotensin Converting Enzyme 1 (gray; Opal™570) on mouse kidney.

Panel B: anti-PTPRO staining glomerulus in mouse kidney.

Panel C: anti-SCL12A1 staining distal tubules in mouse kidney.

Panel D: anti-Angiotensin Converting Enzyme 1 staining proximal tubules in mouse kidney.

Nuclear DNA was labeled with DAPI (shown in blue).
The section was incubated in three rounds of staining: in the order of Anti-GLEPP1/PTPRO antibody [EPR28645-553] ab322047, ab313640 and Anti-Angiotensin Converting Enzyme 1 antibody [EPR22291-247] ab254222 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Nuclear counter stain with DAPI.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: liver, lung, spleen (PMID: 7573490).
The expression profile/ molecular weight observed is consistent with what has been described in literature (PMID: 22759959).
In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
Samples are non-boiled as boiling may cause protein aggregation.
Exposure time: 180 seconds
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640) at 1/1000 dilution
Lane 1: Mouse kidney tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Lane 3: Mouse spleen tissue lysate at 20 µg
Lane 4: Mouse lu tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 150, 270 kDa
Exposure time: 180s
Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody does not cross-react with SLC12A2.

In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640) at 1/1000 dilution
Lane 1: His-tagged mouse SLC12A1/NKCC2 fragment at 2 ng
Lane 2: His-tagged mouse SLC12A2 fragment at 30 ng
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 25 kDa
Exposure time: 180s
Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
SLC12A1/NKCC2 is a glycoprotein of approximately 150 kDa and detected as a 135-kDa band after treated with deglycosylation treatment.

The expression profile/ molecular weight observed is consistent with what has been described in literature (PMID: 22759959).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 15 seconds
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640) at 1/1000 dilution
Lane 1: Untreated Mouse kidney tissue lysate at 20 µg
Lane 2: Mouse kidney tissue lysate treated with Peptide:N-glycosidase F (PNGase F) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 135, 150, 270 kDa
Exposure time: 15s
Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: liver (PMID: 7573490).

The expression profile/ molecular weight observed is consistent with what has been described in literature (PMID: 22759959).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
Exposure time: 48 seconds
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640) at 1/1000 dilution
Lane 1: Rat kidney tissue lysate at 20 µg
Lane 2: Rat liver tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 150 kDa
Exposure time: 48s
Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: liver, lung (PMID: 7573490).

The expression profile/ molecular weight observed is consistent with what has been described in literature (PMID: 22759959).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640) at 1/1000 dilution
Lane 1: Human kidney tissue lysate at 20 µg
Lane 2: Human liver tissue lysate at 20 µg
Lane 3: Human lu tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 150, 270 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on rat liver. The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on mouse liver (PMID: 8621642). The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on human liver. The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the distal tubules of rat kidney. The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the distal tubules of mouse kidney (PMID: 20385770, PMID: 29993276). The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (ab313640)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SLC12A1/NKCC2 with ab313640 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the distal tubules of human kidney. The section was incubated with ab313640 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins