Immunohistochemistry was performed on normal biopsies of deparaffinized Human skeletal muscle tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing PMCA3 ATPase (PA1-916) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
|Tested species reactivity||Human, Pig, Rat|
|Published species reactivity||Pig, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: A(5) N S S I E F H P K P Q Q Q R E V P(22)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1:1,000|
* 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.
PA1-916 detects calcium pump of the plasma membrane 3 (PMCA3 ATPase) from rat, human and porcine tissues.
PA1-916 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~127 kDa protein representing PMCA3a and PMCA3b ATPase from rat brain microsomal fractions.
PA1-916 immunizing peptide corresponds to amino acid residues 5-22 from rat PMCA3 ATPase protein. This sequence is completely conserved between rat and human PMCA3 ATPase.
The calcium pump of the plasma membrane, termed PMCA ATPase, pumps calcium from the cytosol to the extracellular space. This membrane-bound enzyme is related to a number of other ATPases including the SERCA ATPase and the sodium/K+ pump.
There are four different genes encoding PMCA ATPase and studies have revealed 20 isoforms of the pump generated by alternate splicing of the primary gene products. mRNA distribution studies show that gene products 1 and 4 are transcribed in most tissues, however, products 2 and 3 are more tissue specific. Transcription of the splicing variants has also been found to be tissue specific. In the pancreas, where insulin secretion is calcium dependent, the beta cells only express the 4b isoform, however the alpha and gamma cells express both 4a and 4b isoforms. Studies have also shown that different splice variants have different affinities for calcium and calmodulin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Mutation of plasma membrane Ca2+ ATPase isoform 3 in a family with X-linked congenital cerebellar ataxia impairs Ca2+ homeostasis.
PA1-916 was used in immunocytochemistry to study the effects on Ca(2+) homeostasis of a plasma membrane Ca(2+)-ATPase mutation identified in a familiy with X-linked congenital cerebellar ataxia
|Zanni G,Calì T,Kalscheuer VM,Ottolini D,Barresi S,Lebrun N,Montecchi-Palazzi L,Hu H,Chelly J,Bertini E,Brini M,Carafoli E||Proceedings of the National Academy of Sciences of the United States of America (109:14514)||2012|
The plasma membrane Ca2+-ATPase isoform 4 is localized in lipid rafts of cerebellum synaptic plasma membranes.
PA1-916 was used in western blot to localize the plasma membrane calcium-ATPase isoform 4.
|Sepúlveda MR,Berrocal-Carrillo M,Gasset M,Mata AM||The Journal of biological chemistry (281:447)||2006|
Regional distribution of Na,K-ATPase activity in porcine lens epithelium.
PA1-916 was used in western blot to investigate the sodium/potassium ATPase protein expression in porcine lens epithelium.
|Tamiya S,Dean WL,Paterson CA,Delamere NA||Investigative ophthalmology and visual science (44:4395)||2003|