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Immunohistochemistry was performed on cancer biopsies of deparaffinized Human cervical carcinoma 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 PMCA2 ATPase (PA1-915) 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||Rat, Pig, Mouse, Chicken|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: T(5) N S D F Y S K N Q R N E S S(19)|
|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 (Paraffin, Frozen) (IHC (P, F))||1:20-200|
|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.
PA1-915 detects calcium pump of the plasma membrane 2 (PMCA2 ATPase) from human and rat and porcine tissues and cells.
PA1-915 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~127 kDa protein and an ~133 kDa protein representing PMCA2a and PMCA2b ATPase, respectively, from rat brain microsomal fractions.
PA1-915 immunizing peptide corresponds to amino acid residues 5-19 from human PMCA2 ATPase protein. This sequence is completely conserved between human and rat PMCA2 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.
Changes in cochlear PMCA2 expression correlate with the maturation of auditory sensitivity.
PA1-915 was used in western blot to study the relationship between auditory sensitivity maturation and the expression of PMCA2 in a murine model
|Watson CJ,Lies SM,Minich RR,Tempel BL||Journal of the Association for Research in Otolaryngology : JARO (15:543)||2014|
Effects of paraquat-induced oxidative stress on the neuronal plasma membrane Ca(2+)-ATPase.
PA1-915 was used in western blot to study how the intracellular redox cycling agent paraquat affects the functions of PMCA in primary cortical neurons
|Zaidi A,Fernandes D,Bean JL,Michaelis ML||Free radical biology & medicine (47:1507)||2009|
The plasma membrane Ca2+-ATPase isoform 4 is localized in lipid rafts of cerebellum synaptic plasma membranes.
PA1-915 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.
PA1-915 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 & visual science (44:4395)||2003|
A new Atp2b2 deafwaddler allele, dfw(i5), interacts strongly with Cdh23 and other auditory modifiers.
PA1-915 was used in immunohistochemistry to study the interactions of Cdh23 and an unidentified hearing loss gene with a novel Atp2b2 deafwaddler allele of the plasma membrane Ca(2+)-ATPase 2
|Watson CJ,Tempel BL||Hearing research (304:41)||2013|
The development, distribution and density of the plasma membrane calcium ATPase 2 calcium pump in rat cochlear hair cells.
PA1-915 was used in immunohistochemistry to study the localization and function of the plasma membrane calcium ATPase 2 calcium pump in rat cochlear hair cells
|Chen Q,Mahendrasingam S,Tickle JA,Hackney CM,Furness DN,Fettiplace R||The European journal of neuroscience (36:2302)||2012|
PMCA2 via PSD-95 controls calcium signaling by α7-containing nicotinic acetylcholine receptors on aspiny interneurons.
PA1-915 was used in immunocytochemistry to study the mechanism by which plasma membrane calcium-ATPase pump isoform 2 controls calcium signaling in neurons
|Gómez-Varela D,Schmidt M,Schoellerman J,Peters EC,Berg DK||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:6894)||2012|
The novel PMCA2 pump mutation Tommy impairs cytosolic calcium clearance in hair cells and links to deafness in mice.
PA1-915 was used in immunocytochemistry to investigate the effect of the novel PMCA2 pump mutation Tommy for cytosolic calcium clearance in hair cells and deafness in mice
|Bortolozzi M,Brini M,Parkinson N,Crispino G,Scimemi P,De Siati RD,Di Leva F,Parker A,Ortolano S,Arslan E,Brown SD,Carafoli E,Mammano F||The Journal of biological chemistry (285:37693)||2010|
Developmental assembly of transduction apparatus in chick basilar papilla.
PA1-915 was used in immunocytochemistry to study the developmental assembly of the transduction apparatus in chick hair cells.
|Si F,Brodie H,Gillespie PG,Vazquez AE,Yamoah EN||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:10815)||2003|
Interaction of plasma membrane Ca(2+)-ATPase isoform 4 with calcineurin A: implications for catecholamine secretion by PC12 cells.
PA1-915 was used in immunoprecipitation to study the interaction of calcineurin A with PMCA4 and its effect on catecholamine secretion
|Kosiorek M,Podszywalow-Bartnicka P,Zylinska L,Zablocki K,Pikula S||Biochemical and biophysical research communications (411:235)||2011|
ATPase isoform 2 Na+K+ transporting beta polypeptide 2; ATPase isoform 2, Na+K+ transporting, beta polypeptide 2; ATPase, Ca++ transporting, plasma membrane 2; plasma membrane Ca(2+)-ATPase; plasma membrane Ca++-ATPase; plasma membrane Ca2+ pump 2; plasma membrane calcium ATPase; plasma membrane calcium pump; Plasma Membrane Calcium Pump ATPase 2; plasma membrane calcium-transporting ATPase 2
ATP2B2; PMCA2; PMCA2a; PMCA2i