Catalog Fields
Antigen: Na-K-Cl cotransporter
Available to For-Profits: Yes
Antigen Species: Human
Hybridoma Cells Available (Non-Profit): Yes
Isotype: MIgG1
Depositor: Lytle, C. / Forbush III, B.
Host Species: mouse
Antigen Sequence:
Positive Tested Species Reactivity: Mammal
Depositors Institution: Yale University
Antigen Molecular Weight: Predicted: 121.4kDa Apparent: 145-205kDa
Depositors Notes: Antigen retrieval (1% SDS for 5 min) required for immunostaining frozen sections. T9 is better than T4 in recognizing NKCC2, particularly by IF (PMID: 8572179, 12732642). T9 recpgmozes the 196kDa cotransporter exclusively on western blots [PMID: 8910547].
Immunogen: Recombinant 38kDa peptide fragment of carboxy terminus encoded by cDNA from T84 (human) epithelial Na-K-Cl cotransport protein
Gene: SLC12A1, SLC12A2
Alternate Antibody Name:
Alternate Gene Names: NKCC1 and NKCC2
Alternate Antigen Name: NKCC, Solute carrier family 12 members 1 and 2
Clonality: Monoclonal
Myeloma Strain: P3x63-Ag8.653
Epitope Mapped: Yes
Uniprot ID: P55011 Q13621
Epitope Location or Sequence: a.a. 902-1212
Entrez Gene ID: 6557 6558
Immunogen Sequence: human NKCC1 C-terminus (Met902-Ser1212)
Antibody Registry ID: AB_2618107
Recommended Applications: Immunofluorescence, Western Blot
Additional Information: RRID:AB_2618107
All cell products contain the antimicrobial ProClin. Click here for additional information.
These hybridomas were created by your colleagues. Please acknowledge the hybridoma contributor and the Developmental Studies Hybridoma Bank (DSHB) in the Materials and Methods of your publications. Please email the citation to us.
For your Materials & Methods section:
T9 was deposited to the DSHB by Lytle, C. / Forbush III, B. (DSHB Hybridoma Product T9)
Storage and Handling Recommendations
Although many cell products are maintained at 4°C for years without loss of activity, shelf-life at 4°C is highly variable. For immediate use, short term storage at 4°C up to two weeks is recommended. For long term storage, divide the solution into volumes of no less than 20 ul for freezing at -20°C or -80°C. The small volume aliquot should provide sufficient reagent for short term use. Freeze-thaw cycles should be avoided. For concentrate or bioreactor products, an equal volume of glycerol, a cryoprotectant, may be added prior to freezing.
Usage Recommendations
The optimal Ig concentration for an application varies by species and antibody affinity. For each product, the antibody titer must be optimized for every application by the end user laboratory. A good starting concentration for immunohistochemistry (IHC), immunofluorescence (IF), and immunocytochemistry (ICC) when using mouse Ig is 2-5 ug/ml. For western blots, the recommended concentration range of mouse Ig 0.2-0.5 ug/ml. In general, rabbit antibodies demonstrate greater affinity and are used at a magnitude lower Ig concentration for initial testing. The recommended concentrations for rabbit Ig are 0.2-0.5 ug/ml (IF, IHC and ICC) and 20-50 ng/ml (WB).
4 References
Initial Publication
Distribution and diversity of Na-K-Cl cotransport proteins: a study with monoclonal antibodies.
Forbush B 3rd
The American journal of physiology 269.6 Pt 1 (1995 Dec): C1496-505.