Recombinant Anti-NKCC1 antibody [EPR26256-60] (ab303518)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26256-60] to NKCC1
- Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NKCC1 antibody [EPR26256-60]
See all NKCC1 primary antibodies -
Description
Rabbit monoclonal [EPR26256-60] to NKCC1 -
Host species
Rabbit -
Tested applications
Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IHC-Fr -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: His-tagged mouse NKCC1 recombinant protein, human cerebellum, mouse brain, mouse colon cancer, rat brain tissue lysates. IHC-P: Human, mouse, rat colon FFPE tissue sections. ICC/IF: T84 and NIH/3T3 cells. Flow Cyt (Intra): T84 and NIH/3T3 cells. IP: Mouse and rat brain tissue lysate, human cerebellum tissue lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26256-60 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab303518 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IP |
1/30.
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Flow Cyt (Intra) |
1/50.
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WB |
1/1000. Detects a band of approximately 30,150 kDa (predicted molecular weight: 131 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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Notes |
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IP
1/30. |
Flow Cyt (Intra)
1/50. |
WB
1/1000. Detects a band of approximately 30,150 kDa (predicted molecular weight: 131 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Target
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Function
Electrically silent transporter system. Mediates sodium and chloride reabsorption. Plays a vital role in the regulation of ionic balance and cell volume. -
Tissue specificity
Expressed in many tissues. -
Sequence similarities
Belongs to the SLC12A transporter family. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 6558 Human
- Entrez Gene: 20496 Mouse
- Entrez Gene: 83629 Rat
- Omim: 600840 Human
- SwissProt: P55011 Human
- SwissProt: P55012 Mouse
- Unigene: 162585 Human
- Unigene: 712970 Human
see all -
Alternative names
- Basolateral Na-K-Cl symporter antibody
- BSC antibody
- BSC2 antibody
see all
Images
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All lanes : Anti-NKCC1 antibody [EPR26256-60] (ab303518) at 1/1000 dilution
Lane 1 : His-tagged mouse NKCC2 recombinant protein 10 ng
Lane 2 : His-tagged mouse NKCC1 recombinant protein 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 131 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking / diluting buffer and concentration: 5% NFDM/TBST
This antibody does not cross-react with mouse NKCC2.
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All lanes : Anti-NKCC1 antibody [EPR26256-60] (ab303518) at 1/1000 dilution
Lane 1 : Human cerebellum tissue lysate
Lane 2 : Human liver tissue lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : Mouse colon cancer tissue lysate
Lane 5 : Mouse liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 131 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking / diluting buffer and concentration: 5% NFDM/TBST
Low expression: liver (human protein atlas database)
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Anti-NKCC1 antibody [EPR26256-60] (ab303518) + Rat brain tissue lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 131 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Exposure time: 8 secondsBlocking / diluting buffer and concentration: 5% NFDM/TBST
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NKCC1 was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 ug with ab303518 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303518 at dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: mouse brain tissue lysate 10 µg (Inset)
Lane 2: ab303518 IP in mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab303518 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
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NKCC1 was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 ug with ab303518 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303518 at dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: rat brain tissue lysate 10 µg (Inset)
Lane 2: ab303518 IP in rat brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab303518 in rat brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 23 seconds.
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NKCC1 was immunoprecipitated from 0.35 mg human cerebellum tissue lysate 10 ug with ab303518 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303518 at dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: human cerebellum tissue lysate 10 µg (Inset)
Lane 2: ab303518 IP in human cerebellum tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab303518 in human cerebellum tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKCC1 antibody [EPR26256-60] (ab303518)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling NKCC1 with ab303518 at 1/100 dilution (4.89 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND™ Polymer Refine Detection). Positive staining was observed on human colon (PMID: 17478539). The section was incubated with ab303518 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 dilution (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKCC1 antibody [EPR26256-60] (ab303518)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling NKCC1 with ab303518 at 1/2000 dilution (4.89 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND™ Polymer Refine Detection). Positive staining was observed on mouse colon. The section was incubated with ab303518 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 dilution (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKCC1 antibody [EPR26256-60] (ab303518)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling NKCC1 with ab303518 at 1/2000 dilution (4.89 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND™ Polymer Refine Detection). Positive staining was observed on rat colon. The section was incubated with ab303518 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 dilution (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NKCC1 antibody [EPR26256-60] (ab303518)
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling NKCC1 with ab303518 at 1/100 dilution (4.89 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND™ Polymer Refine Detection). Negative control: no staining is observed on human liver. The section was incubated with ab303518 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized T84 (human colon epithelial cell) cells labeling NKCC1 with ab303518 at 1/100 dilution (4.89 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing mainly membranous staining in T84 cell line. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling NKCC1 with ab303518 at 1/100 dilution (4.89 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing membranous and cytoplasmic staining in NIH/3T3 cell line. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized T84 (human colon epithelial cell) cells labeling NKCC1 with ab303518 at 1/50 dilution (1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling NKCC1 with ab303518 at 1/50 dilution (1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab303518 has not yet been referenced specifically in any publications.