Immunofluorescence analysis of IP3 Receptor II was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with IP3 Receptor II Rabbit Polyclonal Antibody (PA1904) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Amphibian, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: E(317) L N P D Y R D A Q N E G K T V R D(334).|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (Frozen) (IHC (F))||1:100|
* 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.
PA1-904 detects inositol 1,4,5-trisphosphate receptor type-II (IP3R-II) from rat and mouse tissues.
PA1-904 has been successfully used in immunohistochemistry, immunofluorescence and immunocytochemistry procedures. Immunohistochemical staining of IP3R-II in rat brain with PA1-904 results in staining of the hippocampus, corpus collosum, and cerebellum. This product fails to detect IP3R-II in Western blot procedures.
PA1-904 immunizing peptide corresponds to amino acid residues 317-334 from rat IP3R-II. PA1-904 immunizing peptide (Cat. # PEP-024) is available for use in neutralization and control experiments.
Inositol 1,4,5-trisphosphate (IP3) is a second messenger for many growth factors, hormones, and neuro-transmitters. Upon binding to IP3R, IP3 triggers the release of intracellular, luminal calcium to the cytosol. Functional IP3R is a homo- or heterotetramer of ~240 kDa glycoprotein subunits. IP3R protein is structurally and functionally related to one other important intracellular calcium-release channel, the ryanodine receptor. The similarity between the two receptors continues at the physiological level owing to a physical association each receptor can have with the immunophilin protein, FKBP12. Phosphorylation of the IP3R by PKC causes an increase in IP3-mediated calcium release. Concomitantly, the phosphatase activity of calcineurin is stimulated upon its association with the FKBP12-IP3R complex. Calcium release is reduced when the PKC target site on the IP3R is dephosphorylated by calcineurin resulting in calcium oscillations. Mammalian IP3R subunits are the product of three distinct genes that are widely expressed and differentially regulated. IP3R type I (IP3R-I) has been detected in heart, liver, kidney, ovary, and Purkinje neurons of the cerebellum. IP3R-II is found predominantly in the brain. IP3R-III is known to be expressed in pancreatic islets, kidney, and the gastrointestinal tract.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Paclitaxel accelerates spontaneous calcium oscillations in cardiomyocytes by interacting with NCS-1 and the InsP3R.
PA1-904 was used in western blot to investigate the effect of taxol on calcium signaling and calcium oscillation
|Zhang K,Heidrich FM,DeGray B,Boehmerle W,Ehrlich BE||Journal of molecular and cellular cardiology (49:829)||2010|
Glycosaminoglycan mimetics trigger IP3-dependent intracellular calcium release in myoblasts.
PA1-904 was used in immunocytochemistry to investigate the effect of glycosaminoglycan mimetics on calcium signaling in myoblasts
|Martelly I,Singabraya D,Vandebrouck A,Papy-Garcia D,Cognard C,Raymond G,Guillet-Deniau I,Courty J,Constantin B||Matrix biology : journal of the International Society for Matrix Biology (29:317)||2010|
Multiple receptor interactions trigger release of membrane and intracellular calcium stores critical for herpes simplex virus entry.
PA1-904 was used in immunocytochemistry to study the role of calcium in herpes simplex viruses entry.
|Cheshenko N,Liu W,Satlin LM,Herold BC||Molecular biology of the cell (18:3119)||2007|
Mini-dystrophin expression down-regulates IP3-mediated calcium release events in resting dystrophin-deficient muscle cells.
PA1-904 was used in immunocytochemistry to study how dystrophin affects IP3-mediaed calcium signaling pathway in resting myotubes
|Balghi H,Sebille S,Mondin L,Cantereau A,Constantin B,Raymond G,Cognard C||The Journal of general physiology (128:219)||2006|
Mini-dystrophin expression down-regulates overactivation of G protein-mediated IP3 signaling pathway in dystrophin-deficient muscle cells.
PA1-904 was used in immunocytochemistry to demonstrate the existence of a potential relationship between mini-dystrophin and SR calcium release
|Balghi H,Sebille S,Constantin B,Patri S,Thoreau V,Mondin L,Mok E,Kitzis A,Raymond G,Cognard C||The Journal of general physiology (127:171)||2006|
Immunolocalization of type 2 inositol 1,4,5-trisphosphate receptors in cardiac myocytes from newborn mice.
PA1-904 was used in immunocytochemistry to investigate the localization of type 2 inositol 1,4,5-trisphosphate receptors (InsP (3) Rs) in cardiac muscle cells
|García KD,Shah T,García J||American journal of physiology. Cell physiology (287:C1048)||2004|
C(a2+)-dependent glutamate release involves two classes of endoplasmic reticulum Ca(2+) stores in astrocytes.
PA1-904 was used in immunocytochemistry to investigate the mechanism of glutamate release induced by calcium in astrocytes
|Hua X,Malarkey EB,Sunjara V,Rosenwald SE,Li WH,Parpura V||Journal of neuroscience research (76:86)||2004|
Immunocytochemical evidence for co-expression of Type III IP3 receptor with signaling components of bitter taste transduction.
PA1-904 was used in immunocytochemistry to study the role of type III IP3 receptor (IP3R3) in bitter taste transduction
|Clapp TR,Stone LM,Margolskee RF,Kinnamon SC||BMC neuroscience (2:null)||2003|
Sparks and puffs in oligodendrocyte progenitors: cross talk between ryanodine receptors and inositol trisphosphate receptors.
PA1-904 was used in immunocytochemistry to investigate the interaction between ryanodine receptors and inositol trisphosphate receptors in oligodendrocyte progenitors.
|Haak LL,Song LS,Molinski TF,Pessah IN,Cheng H,Russell JT||The Journal of neuroscience : the official journal of the Society for Neuroscience (21:3860)||2001|
Expression of calcium transporters in the retina of the tiger salamander (Ambystoma tigrinum).
PA1-904 was used in immunohistochemistry to investigate how cytoplasmic free calcium in neurons from the salamander retina is regulated
|Krizaj D,Liu X,Copenhagen DR||The Journal of comparative neurology (475:463)||2004|
ET-1 activates Ca2+ sparks in PASMC: local Ca2+ signaling between inositol trisphosphate and ryanodine receptors.
PA1-904 was used in immunohistochemistry to suggest that ET-1 activation of calcium sparks is mediated via the ETA receptor-PLC-IP3 pathway
|Zhang WM,Yip KP,Lin MJ,Shimoda LA,Li WH,Sham JS||American journal of physiology. Lung cellular and molecular physiology (285:L680)||2003|