Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling TRPV4 with ab307444 at 1/50 (9.64 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control : no staining on human pancreas. The section was incubated with ab307444 at 4ЎгC overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling TRPV4 with ab307444 at 1/50 (9.64 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human kidney. The section was incubated with ab307444 at 4ЎгC overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

TRPV4 was immunoprecipitated from 0.35 mg A375 (human malignant melanoma epithelial cell) whole cell lysate 10 ug with ab307444 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307444 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : A375 (human malignant melanoma epithelial cell) whole cell lysate 10 ug
Lane 2 : ab307444 IP in A375 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307444 in A375 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 84 seconds

All lanes:

Immunoprecipitation - Anti-TRPV4 antibody [EPR27068-39] (<a href='/en-us/products/primary-antibodies/trpv4-antibody-epr27068-39-ab307444'>ab307444</a>) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate 10 μg

Lane 2:

A375 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 100 kDa

false

Exposure time: 84s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling TRPV4 with ab307444 at 1/1000 (0.482 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on mouse kidney. The section was incubated with ab307444 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND? RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TRPV4 with ab307444 at 1/1000 (0.482 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on choroid plexus of mouse cerebrum. The section was incubated with ab307444 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND? RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeABilized frozen Mouse pancreas (fresh) tissue lABeling TRPV4 with ab307444 at 1/100 (4.82 ug/ml) dilution followed by ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Low expression : confocal image showing no staining on mouse pancreas. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307444 for 60 mins at room temperature. The section was then mounted using Fluoromount?.The immunostaining was performed on a Leica Biosystems BOND? RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using ab307444, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeABilized frozen Mouse kidney (fresh) tissue lABeling TRPV4 with ab307444 at 1/100 (4.82 ug/ml) dilution followed by ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Panel A : merged staining of anti-TRPV4 (ab307444, green) and anti-Nestin (ab306599, red) on mouse kidney.Panel B : anti-TRPV4 stained in the renal tubular system.Panel C : anti-Nestin stained in the glomeruli of kidney.The section was incubated in two rounds of staining : in the order of ab307444 and ab306599 for 1 hr at room temperature. The section was then mounted using Fluoromount?.The immunostaining was performed on a Leica Biosystems BOND? RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbedat 1/1000 2 ug/mL dilution.

• WB

Supplier Data

Western blot - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using 307444, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Negative control : 293T (PMID : 29433476) Positive control : A375 (PMID : 30881453) 70 seconds Exposure time :

All lanes:

Western blot - Anti-TRPV4 antibody [EPR27068-39] (<a href='/en-us/products/primary-antibodies/trpv4-antibody-epr27068-39-ab307444'>ab307444</a>) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate 20 μg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate 20 μg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 100 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-TRPV4 antibody [EPR27068-39] - BSA and Azide free (AB307445)

This data was developed using 307444, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : pancreas (PMID : 14517216)

The MW is consistent to what has been described in the literature (PMID : 30881453)

Samples are non-boiled as boiling may cause protein aggregation.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-TRPV4 antibody [EPR27068-39] (<a href='/en-us/products/primary-antibodies/trpv4-antibody-epr27068-39-ab307444'>ab307444</a>) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human pancreas tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse pancreas tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 100 kDa

true

Exposure time: 180s