Anti-Pannexin 1 antibody [EPR28631-135]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of paraffin-embedded (A) SH-SY5Y (human neuroblastoma epithelial cell) cell pellet, (B) PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) cell pellet and (C) U-937 (human histiocytic lymphoma monocyte) cell pellet labeling Pannexin 1 with ab325529 at 1/5000 (0.098 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive on (A) SH-SY5Y and (B) PANC-1 cell pellet, weak staining on (C) U-937 cell pellet.
The primary antibody was incubated 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Pannexin 1 with ab325529 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human liver.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) and U-937 (human histiocytic lymphoma monocyte) cells labelling Pannexin 1 with ab325529 at 1/200 (2.45 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing membranous staining in PANC-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Low expression : U-937

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab325529 at a 1/200 dilution followed by ab150120 at a 1/1000 dilution.

-ve control 2 : ab7291 at a 1/1000 dilution followed by ab150081 at a 1/1000 dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E18 brain (perfused fixed) tissue labeling Pannexin 1 with ab325529 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Confocal image showing positive staining on mouse E18 brain. The nuclear counterstain was DAPI (Blue). The section was incubated with the primary antibody 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).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (perfused fixed) tissue labeling Pannexin 1 with ab325529 at 1/50 (9.8 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Low expression : confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with the primary antibody 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).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of paraffin-embedded C57BL/6JGpt mouse skeletal muscle tissue labeling Pannexin 1 with ab325529 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on C57BL/6JGpt mouse skeletal muscle.
The primary antibody was incubated 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).

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-Pannexin 1 antibody [EPR28631-135] (AB325529)

Immunohistochemical analysis of paraffin-embedded (A) C57BL/6JGpt mouse E18 brain and (B) C57BL/6JGpt mouse Panx1 KO E18 brain tissue labeling Pannexin 1 with ab325529 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive on (A) C57BL/6JGpt mouse E18 brain and negative on (B) C57BL/6JGpt Panx1 KO mouse E18 brain.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Panx1-KO homozygous mice (Strain ID : T006607).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

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Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

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

Low expression : CT26.WT, U-937.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 20944749, 17925379, 19692571, 25239622).

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

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (124 kDa) staining at 1/10000 dilution.

Exposure time : Lanes 1-2 : 59 seconds, lanes 3-4 : 26 seconds

All lanes:

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (ab325529) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

CT26.WT (mouse colon fibroblast) whole cell lysate at 20 µg

Lane 3:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

U-937 (human histiocytic lymphoma monocyte) whole cell 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 1/20000 dilution

Observed band size: 37-50 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 20944749, 17925379, 19692571, 25239622).

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

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Panx1-KO homozygous mice (Strain ID : T006607).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (ab325529) at 1/1000 dilution

Lane 1:

Wide-type mouse E18 brain tissue lysate at 50 µg

Lane 2:

Panx1 knockout mouse E18 brain tissue lysate at 50 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 1/20000 dilution

Observed band size: 37-50 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (AB325529)

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

Low expression : kidney, liver (PMID : 16026466).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 20944749, 17925379, 19692571, 25239622).

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (ab325529) at 1/1000 dilution

Lane 1:

Mouse olfactory bulb tissue lysate at 50 µg

Lane 2:

Mouse cerebellum tissue lysate at 50 µg

Lane 3:

Mouse kidney tissue lysate at 50 µg

Lane 4:

Mouse liver tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 37-50 kDa,36 kDa

false

Exposure time: 8s