Immunohistochemistry was performed on human renal carcinoma tissue. To expose target protein, antigen was retreived using 10mM sodium citrate followed by microwave treatment for 8-15 minutes. Endogenous peroxidases were blocked in 3% H202-methanol for 15 minutes and tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Sodium/Potassium ATPase mouse monoclonal antibody (Product # MA3-924) at a dilution of 1:10 overnight in a humidified chamber. Tissues were washed in PBST and detection was performed using a secondary antibody conjugated to HRP. DAB staining buffer was applied and tissues were counterstained with hematoxylin and prepped for mounting. Images were taken at 40X magnification.
|Tested species reactivity||Dog, Chicken, Human, Mouse, Rat|
|Published species reactivity||Rat, Pig, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified rat kidney sodium/potassium ATPase.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||1/50|
|Inhibition Assays (IA)||Assay Dependent|
|Immunocytochemistry (ICC)||Assay dependent|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1/100|
|Immunoprecipitation (IP)||Assay dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA3-924 detects the alpha 1 sodium/potassium ATPase in human, mouse, canine, rat and avian tissues.
MA3-924 has been successfully used in immunohistochemistry, immunocytochemistry, immunofluorescence, ELISA and immunoprecipitation procedures. Immunohistochemical staining of sodium/potassium ATPase in rat liver with MA3-924 yields a pattern consistent with plasma membrane localization. Immunoprecipitation of sodium/potassium ATPase from canine kidney extracts can be used for affinity purification of the alpha subunit. MA3-924 can also be utilized to inhibit activity of the enzyme.
The MA3-924 antigen is purified rat kidney sodium/potassium ATPase. This antibody recognizes an epitope within an intracellular region at or near Asp-369 of the sodium/potassium ATPase alpha subunit.
The sodium/potassium ATPase is an integral membrane enzyme found in all cells of higher organisms and is responsible for the ATP-dependent transport of sodium and potassium across the cell membrane. This membrane-bound enzyme is related to a number of other ATPases including sarcoplasmic and endoplasmic reticulum calcium ATPase (SERCA) and plasma membrane calcium ATPase (PMCA). The sodium/potassium ATPase consists of a large, multipass, transmembrane catalytic subunit, termed the alpha subunit, and an associated smaller glycoprotein, termed the beta subunit. Studies indicate that there are three isoforms of the alpha subunit (alpha 1, alpha 2, alpha 3) and two isoforms of the beta subunit (beta 1 and beta 2) encoded by two multigene families.
The different isoforms of the sodium/potassium ATPase exhibit tissue-specific and developmental patterns of expression. The alpha 1 and beta mRNAs are present in all cell types examined, whereas the alpha 2 and alpha 3 mRNAs exhibit a more restricted pattern of cell-specific expression. The alpha subunit has been found in kidney, brain, heart, and to a lesser extent liver, skeletal and smooth muscle.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Characterization of CFTR High Expresser cells in the intestine.
MA3-924 was used in immunohistochemistry to study the function of CFTR High Expresser cells in the intestine of rats
|Jakab RL,Collaco AM,Ameen NA||American journal of physiology. Gastrointestinal and liver physiology (305:G453)||2013|
A membrane-proximal basic domain and cysteine cluster in the C-terminal tail of CCR5 constitute a bipartite motif critical for cell surface expression.
MA3-924 was used in immunohistochemistry to study the mechanism in CCR5 cell surface expression.
|Venkatesan S,Petrovic A,Locati M,Kim YO,Weissman D,Murphy PM||The Journal of biological chemistry (276:40133)||2001|
Hepatic Na(+)-K(+)-ATPase enzyme activity correlates with polarized beta-subunit expression.
MA3-924 was used in immunohistochemistry to investigate the relationship between hepatic sodium/potassium ATPase activity and the expression of polarized beta subunit
|Simon FR,Leffert HL,Ellisman M,Iwahashi M,Deerinck T,Fortune J,Morales D,Dahl R,Sutherland E||The American journal of physiology (269:C69)||1995|
The tetraspanin protein peripherin-2 forms a complex with melanoregulin, a putative membrane fusion regulator.
MA3-924 was used in immunocytochemistry to study the interaction between tetraspanin protein peripherin-2 and melanoregulin
|Boesze-Battaglia K,Song H,Sokolov M,Lillo C,Pankoski-Walker L,Gretzula C,Gallagher B,Rachel RA,Jenkins NA,Copeland NG,Morris F,Jacob J,Yeagle P,Williams DS,Damek-Poprawa M||Biochemistry (46:1256)||2007|
Involvement of IL-10 in exhaustion of myeloid dendritic cells and rescue by CD40 stimulation.
MA3-924 was used in ELISA to study the role of interleukin 10 in the function of myeloid dendritic cells
|Kajino K,Nakamura I,Bamba H,Sawai T,Ogasawara K||Immunology (120:28)||2007|
Identification of monoclonal antibody binding domains of Na+,K(+)-ATPase by immunoelectron microscopy.
MA3-924 was used in crystallagraphy to investigate the binding domains of sodium/potassium ATPase antibody.
|Ting-Beall HP,Beall HC,Hastings DF,Friedman ML,Ball WJ||FEBS letters (265:121)||1990|
Rat hepatic (Na+, K+)-ATPase: alpha-subunit isolation by immunoaffinity chromatography and structural analysis by peptide mapping.
MA3-924 was used in neutralization experiments in rat liver
|Hubert JJ,Schenk DB,Skelly H,Leffert HL||Biochemistry (25:4156)||1986|
Use of a monoclonal antibody to quantify (Na+,K+)-ATPase activity and sites in normal and regenerating rat liver.
MA3-924 was used in blocking/activating experiment to investigate the activity and sites of sodium/potassium ATPase in normal and regenerating rat liver
|Schenk DB,Hubert JJ,Leffert HL||The Journal of biological chemistry (259:14941)||1984|