Anti-TRPV4 antibody (ab39260)
Key features and details
- Rabbit polyclonal to TRPV4
- Suitable for: ICC, WB, IHC-P
- Reacts with: Mouse, Horse
- Isotype: IgG
Overview
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Product name
Anti-TRPV4 antibody
See all TRPV4 primary antibodies -
Description
Rabbit polyclonal to TRPV4 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC MouseIHC-P MouseWB Mouse
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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
- ICC: TKPTS cells
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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Immunocytochemistry - Anti-TRPV4 antibody (ab39260) Lab
ab39260 staining TRPV4 in TKPTS cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab39260 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody (ab39260)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.
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Western blot - Anti-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
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody (ab39260) This image is courtesy of an Abreview submitted by Mr Carl Hobbsab39260 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. -
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
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.
