Anti-NKCC1/SLC12A2 Antibody (monoclonal, 6G7D2)

• Catalog Number: orb1184753
• Category: Antibodies
• Description: Anti-NKCC1/SLC12A2 Antibody (monoclonal, 6G7D2). Tested in IF, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 6G7D2
• Tested applications: IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Mouse IgG2a
• Immunogen: E.coli-derived human NKCC1/SLC12A2 recombinant protein (Position: K889-K943).
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 200 kDa
• UniProt ID: P55011
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Eukaryotic translation initiation factor 6; eIF-6;
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human, Mouse, Rat Immunofluorescence, 5 μg/ml, Human, Mouse, Rat. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Biotin conjugated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of mouse colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Biotin conjugated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of rat colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Biotin conjugated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of mouse colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. NKCC1/SLC12A2 was detected in a paraffin-embedded section of rat colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-NKCC1/SLC12A2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A549 whole cell lysates, Lane 2: human CACO-2 whole cell lysates, Lane 3: human PC-3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-NKCC1/SLC12A2 antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NKCC1/SLC12A2 at approximately 200 kDa. The expected band size for NKCC1/SLC12A2 is at 131 kDa.