This antibody gave a positive signal in Mouse Brain tissue lysate.
Although some of our customers have had good results in Human and Rat (89% homology with the immunogen), we do not batch test this antibody in these species. Due to the polyclonal nature of this antibody, some batches many not work in Human or Rat.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 98 kDa).
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Non-selective calcium permeant cation channel probably involved in osmotic sensitivity and mechanosensitivity. Activation by exposure to hypotonicity within the physiological range exhibits an outward rectification. Also activated by low pH, citrate and phorbol esters. Increase of intracellular Ca(2+) potentiates currents. Channel activity seems to be regulated by a calmodulin-dependent mechanism with a negative feedback mechanism. Promotes cell-cell junction formation in skin keratinocytes and plays an important role in the formation and/or maintenance of functional intercellular barriers. Acts as a regulator of intracellular Ca(2+) in synoviocytes. Plays an obligatory role as a molecular component in the nonselective cation channel activation induced by 4-alpha-phorbol 12,13-didecanoate and hypotonic stimulation in synoviocytes and also regulates production of IL-8.
Found in the synoviocytes from patients with (RA) and without (CTR) rheumatoid arthritis (at protein level).
Brachyolmia 3 Spondylometaphyseal dysplasia Kozlowski type Metatropic dysplasia Distal spinal muscular atrophy, congenital non-progressive Charcot-Marie-Tooth disease 2C Scapuloperoneal spinal muscular atrophy Spondyloepiphyseal dysplasia Maroteaux type Parastremmatic dwarfism Digital arthropathy-brachydactyly, familial
Belongs to the transient receptor (TC 1.A.4) family. TrpV subfamily. TRPV4 sub-subfamily. Contains 3 ANK repeats.
Phosphorylation results in enhancement of its channel function.
Cell membrane and Cell membrane. Cell junction > adherens junction. Assembly of the putative homotetramer occurs primarily in the endoplasmic reticulum.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - TRPV4 antibody (ab39260)This image is courtesy of an Abreview submitted by Mr Carl Hobbs
ab39260 staining of TRPV4 -/- (upper panel) and wildtype (lower panel) adult mouse kidney tissue sections, showing good immunostaining in the wildtype tissue and no immunostaining in the TRPV4 -/- tissue. Formalin/PFA-fixed paraffin-embedded sections of mouse kidney tissue were incubated with ab39260 (1/200) for 2 hours. Antigen retrieval was performed by heat induction in citrate buffer pH 6.0. A biotin-conjugated goat anti-rabbit antibody was used as the secondary.
ICC/IF image of ab39260 stained PC12 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab39260, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - TRPV4 antibody (ab39260)
Anti-TRPV4 antibody (ab39260) at 2 µg/ml + Brain (Mouse) Tissue Lysate at 25 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Immunohistochemistry (PFA perfusion fixed frozen sections) - TRPV4 antibody (ab39260)This image is courtesy of an Abreview submitted by Sophie Pezet, ESPCI, France
IHC-FoFr image of TRPV4 staining on Rat DRG sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the tissues were cryoprotected in sucrose 30% overnight. Tissues were then cut using a cryostat. The image shows DRG neurons (left), nerve taken at the level of the root (right)
Western blot - Anti-TRPV4 antibody (ab39260)Image courtesy of an anonymous Abreview.
All lanes : Anti-TRPV4 antibody (ab39260) at 1 µg/ml
Lane 1 : Whole cell lysate prepared from MEF cells Lane 2 : Whole cell lysate prepared from F9 cells Lane 3 : Whole cell lysate prepared from GC-1 cells Lane 4 : Whole cell lysate prepared from NIH-3T3 cells
Lysates/proteins at 20 µg per lane.
Secondary All lanes : HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution
This antibody detects a band of ~125 kDa in 4 different mouse cell lines. This is lower than the predicted molecular weight of TRPV4 on SDS-PAGE (100 kDa). However, there are published reports of TRPV4 running at >120 kDa PMID: 16368742 PMID: 12538589, possibly as a result of glycosylation.