Anti-TRPV4 antibody

• IHC-P

Lab

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 Bond^TM 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TRPV4 antibody (AB39260)

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.

• IHC-P

AbReview10373****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody (AB39260)

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.

This image is courtesy of an Abreview submitted by Mr Carl Hobbs

• WB

Ap

Western blot - Anti-TRPV4 antibody (AB39260)

All lanes:

Western blot - Anti-TRPV4 antibody (ab39260) at 2 µg/mL

All lanes:

Brain (Mouse) Tissue Lysate at 25 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 98 kDa

Observed band size: 100 kDa,37 kDa,54 kDa

true

Exposure time: 1min

• WB

AbReview28296****

Western blot - Anti-TRPV4 antibody (AB39260)

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.

All lanes:

Western blot - Anti-TRPV4 antibody (ab39260) at 1 µg/mL

Lane 1:

Whole cell lysate prepared from MEF cells at 20 µg

Lane 2:

Whole cell lysate prepared from F9 cells at 20 µg

Lane 3:

Whole cell lysate prepared from GC-1 cells at 20 µg

Lane 4:

Whole cell lysate prepared from NIH-3T3 cells at 20 µg

Secondary

All lanes:

HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution

Predicted band size: 98 kDa

Observed band size: 125 kDa

true

Exposure time: 5min

Image courtesy of an anonymous Abreview.