Anti-Glypican 1/ GPC1 antibody [EPR30349-529]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse pancreatic carcinoma tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreatic carcinoma. 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-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse squamous cell carcinoma tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse squamous cell carcinoma. 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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized B16-F10 (mouse skin melanoma cell) cells labelling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in B16-F10 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : A20.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ab150081 1000 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on scattered cells of mouse liver (PMID : 32228854). 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-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum (PMID : 32228854). 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-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cardiac muscle (PMID : 32228854). 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

• I-ELISA

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Indirect ELISA - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Indirect ELISA analysis of ab324685 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Mouse Glypican-1, Mouse Glypican-6.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Mouse Glypican-6 by ELISA.

• WB

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Western blot - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

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

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

All lanes:

Western blot - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (ab324685) at 1/1000 dilution

Lane 1:

B16-F10 (mouse skin melanoma cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

B16-F10 transfected with siRNA specifically targeting GPC1 whole cell lysate at 40 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: 61 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

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

Low expression : A20. Mouse lung, liver(PMID : 32228854, PMID : 9438425).

The molecular weight observed is consistent with what has been described in the literature PMID : 22351761.

LMW : Core GPC1 protein

HMW : Heparan sulfate substitution on the core GPC1 protein

Blot of lane 1 and lane 2 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range. Remaining wells were developed using the standard substrate protocol.

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

All lanes:

Western blot - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (ab324685) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse skin tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse liver tissue lysate at 20 µg

Lane 6:

Mouse lung tissue lysate at 20 µg

Lane 7:

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

Lane 8:

A20 (mouse reticulum sarcoma B lymphocyte) 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 1/100000 dilution

Observed band size: 61-300 kDa,36 kDa

true

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 1/ GPC1 antibody [EPR30349-529] (AB324685)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Glypican 1/ GPC1 with ab324685 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : almost no staining on mouse pancreas. 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