Anti-GPCR GPR97 antibody [EPR30227-586]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling GPCR GPR97 with ab325332 at 1/100 (5.18 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Positive staining on granulocytes of human placenta.

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-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling GPCR GPR97 with ab325332 at 1/100 (5.18 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Positive staining on human spleen.

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-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling GPCR GPR97 with ab325332 at 1/100 (5.18 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Positive staining on granulocytes of 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.

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-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Immunohistochemical analysis of paraffin-embedded :
(A) SU-DHL-1 (human large cell lymphoma histiocytic cell) cell pellet
(B) MCF7 (human breast adenocarcinoma epithelial cell) cell pellet
(C) HeLa (human cervical adenocarcinoma epithelial cell) cell pellet
labeling GPCR GPR97 with ab325332 at 1/500 (1.036 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Positive staining on (A) : SU-DHL-1(human large cell lymphoma histiocytic cell) cell pellet.

No staining on (B) : MCF7 (human breast adenocarcinoma epithelial cell) cell pellet, and (C) : HeLa (human cervical adenocarcinoma epithelial cell) 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-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Immunohistochemical analysis of paraffin-embedded :
(A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human AGRG3/GPR97 expression vector containing a Myc-His tag cell pellet.
(B) HEK-293T cells transfected with empty vector containing a myc-His-tag.
labeling GPCR GPR97 with ab325332 at 1/100 (5.18 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Positive staining on (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human AGRG3/GPR97 expression vector containing a myc-His-tag. No staining on (B) HEK-293T cells transfected with empty vector containing a myc-His-tag.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining GPCR GPR97 with ab325332 at a 1 : 100 (4.97 µg/ml) dilution, ab238794 anti-CD11b used at 1 : 400 (2.9 µg/ml) dilution.

Panel A : merged staining of anti-GPCR GPR97 (green; Opal™520) and anti-CD11b (magenta; Opal™570) on human placenta.
Panel B : anti-GPCR GPR97 staining granulocytes in human placenta.
Panel C : ant-CD11b staining granulocytes in human placenta.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325332 and ab238794 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GPCR GPR97 antibody [EPR30227-586] (AB325332)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining GPCR GPR97 with ab325332 at a 1 : 100 (4.97 µg/ml) dilution, ab238794 anti-CD11b used at 1 : 400 (2.9 µg/ml) dilution.

Panel A : merged staining of anti-GPCR GPR97 (green; Opal™520) and anti-CD11b (magenta; Opal™570) on human liver.
Panel B : anti-GPCR GPR97 staining granulocytes in human liver.
Panel C : ant-CD11b staining granulocytes in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325332 and ab238794 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-GPCR GPR97 antibody [EPR30227-586] (AB325332)

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

Low expression : MCF7, HeLa

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

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda).

All lanes:

Western blot - Anti-GPCR GPR97 antibody [EPR30227-586] (ab325332) at 1/1000 dilution

Lane 1:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate at 40 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) 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 1/20000 dilution

Observed band size: 38 kDa,15 kDa

false

Exposure time: 180s