Anti-EDG2/LPA-1 antibody [EPR30753-555]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human endometrium (PMID : 18617617, PMID : 29328374). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded (A) : MG-63 (human osteosarcoma fibroblast cell) cell pellet (B) : HepG2 (human hepatocellular carcinoma epithelial cell) cell pellet tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive on (A) : MG-63 cell pellet, no staining on (B) : HepG2 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human glioma. 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on stroma in human liver (PMID : 9070858). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on stroma in rat 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.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on stroma in mosue 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.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum (PMID : 11948806). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling EDG2 /LPA-1 with ab325552 at 1/500 (1.028 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum. 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

• WB

Lab

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

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

Low expression : liver(PMID : 9070858)

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

Lane1 is incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 2-3 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (ab325552) at 1/1000 dilution

Lane 1:

Human breast tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Secondary

Lane 1:

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

Lanes 2 - 3:

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: 25-75 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

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

Low expression : HepG2

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-4 : 37 seconds, lanes 5-6 : 15 seconds

All lanes:

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (ab325552) at 1/1000 dilution

Lane 1:

U-118 MG (human brain glioblastoma cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

U-118 MG transfected with siRNA specifically targeting LPAR1 whole cell lysate at 20 µg

Lane 3:

Untreated U-118 MG whole cell lysate at 20 µg

Lane 4:

U-118 MG whole cell lysate treated with Protein Deglycosylation Peptide:N-glycosidase F (PNGase F) at 20 µg

Lane 5:

MG-63 (human osteosarcoma fibroblast) whole cell lysate at 20 µg

Lane 6:

HepG2 (human hepatocellular carcinoma epithelial cell) 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: 25-75 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (AB325552)

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

Low expression : liver(PMID : 9070858)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-2 : 37 seconds, lane 3 : 81 seconds

All lanes:

Western blot - Anti-EDG2/LPA-1 antibody [EPR30753-555] (ab325552) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

C6 (rat glial tumor glial cell) 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: 25-75 kDa,36 kDa

false