SLC12A2 / NKCC1 Recombinant Rabbit Monoclonal Antibody [PSH23-65] - BSA and Azide free
Catalog# HA751906
SLC12A2 / NKCC1 Recombinant Rabbit Monoclonal Antibody [PSH23-65] - BSA and Azide free
-
WB
-
IF-Cell
-
IHC-Fr
-
IHC-P
-
FC
-
IP
-
Human
-
Mouse
-
Rat
-
HA724390
含抗保成分
-
unconjugated
概述
产品名称
SLC12A2 / NKCC1 Recombinant Rabbit Monoclonal Antibody [PSH23-65] - BSA and Azide free
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Recombinant protein within mouse SLC12A2 / NKCC1 aa 101-279.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IHC-Fr, IHC-P, FC, IP
分子量
Predicted band size: 131 kDa
阳性对照
COLO205 (Human colon adenocarcinoma cells) cell lysate, HT-29 (Human colorectal adenocarcinoma cells) cell lysate, HeLa (Human cervical adenocarcinoma cells) cell lysate, NIH/3T3 (Mouse fibroblasts) cell lysate, Mouse cerebellum tissue lysate, Mouse brain tissue lysate, Mouse colon tissue lysate, Mouse small intestine tissue lysate, Rat cerebellum tissue lysate, Rat brain tissue lysate.
偶联
unconjugated
克隆号
PSH23-65
产品特性
形态
Liquid
浓度
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
-
WB
-
1:5,000
-
IF-Cell
-
1:500
-
IHC-Fr
-
1:500
-
IHC-P
-
1:5,000
-
FC
-
1:1,000
-
IP
-
1-2μg/sample
靶点
功能
The Na–K–Cl cotransporter (NKCC) is a transport protein that aids in the secondary active transport of sodium, potassium, and chloride into cells. In humans there are two isoforms of this membrane transport protein, NKCC1 and NKCC2, encoded by two different genes (SLC12A2 and SLC12A1 respectively). Two isoforms of the NKCC1/Slc12a2 gene result from keeping (isoform 1) or skipping (isoform 2) exon 21 in the final gene product. NKCC1 is widely distributed throughout the human body; it has important functions in organs that secrete fluids. In contrast, NKCC2 is found specifically in the kidney, where it extracts sodium, potassium, and chloride from the urine so they can be reabsorbed into the blood.
背景文献
1. Zhou J et al. Exosome-derived SLC12A2-DT promotes macrophage M2 polarization-mediated hepatocellular carcinoma progression. Int Immunopharmacol. 2025 Jul
2. Delpire E et al. NKCC1 in human diseases: is the SLC12A2 gene haploinsufficient? Am J Physiol Cell Physiol. 2023 Aug
亚细胞定位
Basolateral cell membrane.
别名
Solute carrier family 12 member 2
Basolateral Na-K-Cl symporter
Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2 (BSC2)
Na-K-2Cl cotransporter 1 (hNKCC1)
SLC12A2
NKCC1
图片
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Western blot analysis of SLC12A2 / NKCC1 on different lysates with Rabbit anti-SLC12A2 / NKCC1 antibody (HA751906) at 1/5,000 dilution.
Lane 1: COLO205 (Human colon adenocarcinoma cells) cell lysate
Lane 2: HT-29 (Human colorectal adenocarcinoma cells) cell lysate
Lane 3: HeLa (Human cervical adenocarcinoma cells) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblasts) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 20 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751906, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 131 kDa
Observed band size: 150-200 kDa -
☑ Relative expression (RE)
Western blot analysis of SLC12A2 / NKCC1 on different lysates with Rabbit anti-SLC12A2 / NKCC1 antibody (HA751906) at 1/5,000 dilution.
Lane 1: Mouse cerebellum tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Mouse liver tissue lysate
Lane 4: Mouse colon tissue lysate
Lane 5: Mouse small intestine tissue lysate
Lane 6: Rat cerebellum tissue lysate
Lane 7: Rat liver tissue lysate
Lane 8: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 20 seconds; ECL: K1801
Low expression: Liver (PMID:12000719)
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751906, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 131 kDa
Observed band size: 150-200 kDa -
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: HT-29 (Human colorectal adenocarcinoma cells)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751906, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue). -
Application: Immunofluorescence (IHC-Fr)
Species: Mouse
Tissue: Colon
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA751906, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunofluorescence (IHC-Fr)
Species: Rat
Tissue: Colon
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA751906, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Cerebellum
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Cerebellum
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
☑ Relative expression (RE)
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Liver
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Negative expression of SLC12A2 / NKCC1 protein in liver is consistent with the predicted expression pattern. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Cerebellum
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751906, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Flow Cytometry (Intra)
Species: Human
Sample: HT-29 (Human colorectal adenocarcinoma cells)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751906 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. -
Immunoprecipitation (IP)
SLC12A2 / NKCC1 was immunoprecipitated in 0.2 mg COLO205 (Human colon adenocarcinoma cells) cell lysate with HA751906 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751906 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: COLO205 cell lysate (input)
Lane 2: HA751906 IP in COLO205 cell lysate
Lane 3: Rabbit IgG instead of HA751906 in COLO205 cell lysate
Exposure time: 7 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA751906, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 131 kDa
Observed band size: 150-200 kDa
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