Overview

  • Product name

  • Description

    Rabbit polyclonal to TRPV4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-FoFr, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Horse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 850 to the C-terminus of Mouse TRPV4.

    Read Abcam's proprietary immunogen policy (Peptide available as ab39471.)

  • Positive control

    • This antibody gave a positive signal in Mouse Brain tissue lysate. This antibody gave a positive signal in Mouse Brain FFPE tissue.
  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab39260 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
IHC-FoFr 1/300.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 98 kDa).
IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    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.
  • Tissue specificity

    Found in the synoviocytes from patients with (RA) and without (CTR) rheumatoid arthritis (at protein level).
  • Involvement in disease

    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
  • Sequence similarities

    Belongs to the transient receptor (TC 1.A.4) family. TrpV subfamily. TRPV4 sub-subfamily.
    Contains 3 ANK repeats.
  • Post-translational
    modifications

    Phosphorylation results in enhancement of its channel function.
  • Cellular localization

    Cell membrane and Cell membrane. Cell junction > adherens junction. Assembly of the putative homotetramer occurs primarily in the endoplasmic reticulum.
  • Information by UniProt
  • Database links

  • Alternative names

    • BCYM3 antibody
    • CMT2C antibody
    • HMSN2C antibody
    • osm 9 like TRP channel 4 antibody
    • Osm-9-like TRP channel 4 antibody
    • OSM9 like transient receptor potential channel 4 antibody
    • Osmosensitive transient receptor potential channel 4 antibody
    • OTRPC 4 antibody
    • OTRPC4 antibody
    • SMAL antibody
    • SPSMA antibody
    • SSQTL1 antibody
    • Transient receptor potential cation channel subfamily V member 4 antibody
    • Transient receptor potential protein 12 antibody
    • TRP 12 antibody
    • TRP12 antibody
    • TRPV 4 antibody
    • TrpV4 antibody
    • TRPV4_HUMAN antibody
    • Vanilloid receptor like channel 2 antibody
    • Vanilloid receptor like protein 2 antibody
    • Vanilloid receptor related osmotically activated channel antibody
    • Vanilloid receptor-like channel 2 antibody
    • Vanilloid receptor-like protein 2 antibody
    • Vanilloid receptor-related osmotically-activated channel antibody
    • VR 4 antibody
    • VR OAC antibody
    • VR-OAC antibody
    • VR4 antibody
    • VRL 2 antibody
    • VRL-2 antibody
    • VRL2 antibody
    • VROAC antibody
    see all

Images

  • IHC image of ab39260 staining in mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) for 20 mins. The section was then incubated with ab39260, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

  • 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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 98 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa, 54 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute
  • 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.

    See Abreview

  • 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).
  • 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)

    See 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

    Developed using the ECL technique.

    Predicted band size: 98 kDa
    Observed band size: 125 kDa why is the actual band size different from the predicted?


    Exposure time: 5 minutes


    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.

    See Abreview

References

This product has been referenced in:

  • Minami A  et al. Hyperbaric oxygen significantly improves frequent urination, hyperalgesia, and tissue damage in a mouse long-lasting cystitis model induced by an intravesical instillation of hydrogen peroxide. Neurourol Urodyn 38:97-106 (2019). Read more (PubMed: 30411813) »
  • Lee HP  et al. Volume expansion and TRPV4 activation regulate stem cell fate in three-dimensional microenvironments. Nat Commun 10:529 (2019). Read more (PubMed: 30705265) »
See all 33 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Application
Western blot
Sample
Pig Cell lysate - whole cell (Ocular cell)
Gel Running Conditions
Reduced Denaturing (4-12)
Loading amount
20 µg
Specification
Ocular cell
Blocking step
LICOR blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 08 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Permeabilization
Yes - Wash Buffer from Dako with Tween
Specification
kidney
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

Submitted May 11 2016

Application
Flow Cytometry
Sample
Human Cell (Aortic endothelial cell)
Permeabilization
No
Gating Strategy
size and granularity
Specification
Aortic endothelial cell
Preparation
Cell harvesting/tissue preparation method: cells were detached from tissue culture plates with 0.05% Trypsin-EDTA
Sample buffer: 2mM EDTA + 0.5% BSA in PBS
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 30 2015

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (Lung)
Specification
Lung
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 05 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Dako Flex peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH
Sample
Human Tissue sections (Tendon)
Specification
Tendon
Permeabilization
No
Fixative
10% buffered formalin

Abcam user community

Verified customer

Submitted Oct 23 2014

Application
Western blot
Loading amount
200 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10% gel)
Sample
Rat Tissue lysate - whole (bladder)
Specification
bladder
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 18 2013

Answer

Thank you for contacting us. The first image on our website did show a band at ˜ 100 kD as predicted.
There is also a different image from a customer Abreview with a band ˜125 kDa in 4 different mouse cell lines. However, there are published reports of TRPV4 running at >120 kDa PMID: 16368742 PMID: 12538589, possibly as a result of glycosylation.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for contacting us. For blocking we typically test our antibodies with 3% BSA although you may use 5% nonfat milk if that is your preference.

Regarding the concentration for WB:

ab11370 use at 1/8000

ab39262 use 1-2 ug/ml

ab83737 use 1/300-1/2000

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Question
Answer

Thank you very much for your call today and for letting us know about the problem with the vial of this antibody.

I'm sending a free of charge replacement vial on the order ***, which should arrive tomorrow. Please keep me updated about the results with this replacement antibody, and let me know if there is anything else that we can do for you.

Have a nice day.

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse cell lines)
Loading amount
20 µg
Specification
Mouse cell lines
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Feb 23 2012

1-10 of 19 Abreviews or Q&A

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