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 PMCA1 ATPase (PA1-914) 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, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues A(5) N N S V A Y S G V K N S I K E A N(22) of rat PMCA1 ATPase.|
|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.
PA1-914 detects calcium pump of the plasma membrane 1a (PMCA1a ATPase) and 1b from rat, human and porcine tissues.
PA1-914 has been successfully used in Western blot, immunohistochemistry (paraffin and frozen), and ELISA procedures. By Western blot, this antibody detects ~134 kDa and ~130 kDa proteins representing PMCA1a and 1b ATPase from rat, human and porcine samples.
The PA1-914 immunogen is a synthetic peptide corresponding to the residues A(5) N N S V A Y S G V K N S I K E A N(22) of rat PMCA1 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/potassium 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.
Gain-of-function nature of Cav1.4 L-type calcium channels alters firing properties of mouse retinal ganglion cells.
PA1-914 was used in immunohistochemistry to study alteration of firing properties of mouse retinal ganglion cells due to the gain-of-function nature of Cav1.4 L-type calcium channels
|Knoflach D,Schicker K,Glösmann M,Koschak A||Channels (Austin, Tex.) (9:298)||2015|
Genetic control of circuit function: Vsx1 and Irx5 transcription factors regulate contrast adaptation in the mouse retina.
PA1-914 was used in immunohistochemistry to study the function of Vsx1 and Irx5 in the mouse retina.
|Kerschensteiner D,Liu H,Cheng CW,Demas J,Cheng SH,Hui CC,Chow RL,Wong RO||The Journal of neuroscience : the official journal of the Society for Neuroscience (28:2342)||2008|
Histone deacetylase inhibitor- and PMA-induced upregulation of PMCA4b enhances Ca2+ clearance from MCF-7 breast cancer cells.
PA1-914 was used in western blot to study increased breast cancer cell Ca(2+) clearance by PMCA4b upregulated in response to HDAC inhibitors or PMA
|Varga K,Pászty K,Padányi R,Heged¿s L,Brouland JP,Papp B,Enyedi A||Cell calcium (55:78)||2014|
Effect of silencing VDR gene in kidney on renal epithelial calcium transporter proteins and urinary calcium excretion in genetic hypercalciuric stone-forming rats.
PA1-914 was used in western blot to study the role of vitamin D receptor in renal calcium reabsorption
|Xi QL,Wang SG,Ye ZQ,Zhu ZW,Li C,Bai J,Yu X,Liu JH||Urology (78:1442.e1)||2011|
Adenovirus-delivered microRNA targeting the vitamin D receptor reduces intracellular Ca²¿ concentrations by regulating the expression of Ca²¿-transport proteins in renal epithelial cells.
PA1-914 was used in western blot to investigate the influence of vitamin D receptor knockdown on specific gene expression and calcium signal
|Xi Q,Wang S,Ye Z,Liu J,Yu X,Zhu Z,Su S,Bai J,Li C||BJU international (107:1314)||2011|
The plasma membrane Ca2+-ATPase isoform 4 is localized in lipid rafts of cerebellum synaptic plasma membranes.
PA1-914 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.
PA1-914 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|