NKCC1/SLC12A2 Polyclonal antibody

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  WB analysis using 28055-1-AP

  Boiled and unboiled HeLa cell lysates were subjected to SDS PAGE followed by western blot with 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 28055-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:600 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 28055-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:600 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 28055-1-AP

  Immunohistochemical analysis of paraffin-embedded colon cancer slide using 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human prostate cancer using 28055-1-AP

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:600 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human prostate cancer using 28055-1-AP

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 28055-1-AP (NKCC1/SLC12A2 antibody) at dilution of 1:600 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 28055-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using NKCC1/SLC12A2 antibody (28055-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of A549 using 28055-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed A549 cells using NKCC1,SLC12A2 antibody (28055-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

NKCC1/SLC12A2 Polyclonal Antibody for WB, IHC, IF/ICC, ELISA

Cat No. 28055-1-AP

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

NKCC1,SLC12A2, NKCC1, SLC12A2, Basolateral Na K Cl symporter, Basolateral Na-K-Cl symporter

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Tested Applications

• Positive WB detected in: unboiled HeLa cells
• Positive IHC detected in: human breast cancer tissue, human colon cancer tissue, human prostate cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, A549 cells

For optimal WB detection with 28055-1-AP, we do not recommend boiling the sample after lysis.

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:300-1:1200
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

28055-1-AP targets NKCC1/SLC12A2 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag27895

  Product name: Recombinant human SLC12A2 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 1-280 aa of BC033003

  Sequence: MEPRPTAPSSGAPGLAGVGETPSAAALAAARVELPGTAVPSVPEDAAPASRDGGGVRDEGPAAAGDGLGRPLGPTPSQSRFQVDLVSENAGRAAAAAAAAAAAAAAAGAGAGAKQTPADGEASGESEPAKGSEEAKGRFRVNFVDPAASSSAEDSLSDAAGVGVDGPNVSFQNGGDTVLSEGSSLHSGGGGGSGHHQHYYYDTHTNTYYLRTFGHNTMDAVPRIDHYRHTAAQLGEKLLRPSLAELHDELEKEPFEDGFANGEESTPTRDAVVTYTAESK

• Full Name: solute carrier family 12 (sodium/potassium/chloride transporters), member 2
• Calculated Molecular Weight: 131 kDa
• Observed Molecular Weight: 150-160 kDa
• GenBank Accession Number: BC033003
• Gene Symbol: NKCC1/SLC12A2
• Gene ID (NCBI): 6558
• RRID: AB_2881048
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P55011
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The Na-K-Cl cotransporter sodium-potassium-chloride cotransporter 1 (NKCC1), encoded by the SLC12A2 gene, is an intensively studied member of the CCC family. NKCC1 is a membrane protein and plays fundamental roles in regulating trans-epithelial ion movement, cell volume, chloride homeostasis and neuronal excitability. NKCC1 assembles into a dimer, with the first ten transmembrane (TM) helices harboring the transport core and TM11-TM12 helices lining the dimer interface(PMID: 32081947). The observed molecular weight of NKCC1 is about 140 kDa, glycosylated NKCC1 is detected a band at ∼170 kDa (PMID: 22723696).