Anti-BMP2 antibody [EPR30246-26]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling BMP2 with ab325708 at 1/100 (5.2 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver cancer. 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-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded (A) HeLa (human cervical adenocarcinoma epithelial cell) cell pellet and (B) Daudi (human Burkitt's lymphoma lymphoblast) cell pellet tissue labeling BMP2 with ab325708 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HeLa cell pellet, no staining on (B) Daudi 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling BMP2 with ab325708 at 1/100 (5.2 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive 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.

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-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa (human cervical adenocarcinoma epithelial cell) cell pellet and (B) BMP2 knockout HeLa cell pellet labeling BMP2 with ab325708 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild-type HeLa cell pellet, no staining on (B) BMP2 knockout HeLa 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling BMP2 with ab325708 at 1/100 (5.2 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas (PMID : 20959140). 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-Fr

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Immunohistochemistry (Frozen sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (perfused fixed) tissue labeling BMP2 with ab325708 at 1/50 (10.4 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive 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 (2 ug/mL) 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-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling BMP2 with ab325708 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling BMP2 with ab325708 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive 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.

• WB

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Western blot - Anti-BMP2 antibody [EPR30246-26] (AB325708)

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

Low expression : Daudi

The identity of the lower MW band at approximately 12 kDa is unknown.

Lanes 1-5 were incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and Lanes 6-7 were incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

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

Exposure time : Lanes 1-5 : 136 seconds; Lanes 6-7 : 5 seconds

All lanes:

Western blot - Anti-BMP2 antibody [EPR30246-26] (ab325708) at 1/1000 dilution

Lane 1:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Human testis tissue lysate at 20 µg

Lane 6:

Human liver tissue lysate at 20 µg

Lane 7:

Mouse liver tissue lysate at 20 µg

Secondary

Lanes 1 - 5:

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

Lanes 5 - 6:

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

Observed band size: 35 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-BMP2 antibody [EPR30246-26] (AB325708)

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

Performed under reducing conditions.

In Western blot, ab325708 was shown to bind specifically to BMP2. Target of interest was observed at 35 kDa in HeLa cell lysates (lane 1) with no signal observed at this size in BMP2 knockout cell line (lane 2, knockout cell line ab261767 / knockout lysate ab256852).

The identity of the lower MW band at approximately 12 kDa is unknown.

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

All lanes:

Western blot - Anti-BMP2 antibody [EPR30246-26] (ab325708) at 1/1000 dilution

Lane 1:

Western blot - Human BMP2 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-bmp2-knockout-hela-cell-lysate-ab256852'>ab256852</a>) at 20 µg

Lane 2:

BMP2 knockout HeLa 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: 35 kDa,36 kDa

false

Exposure time: 70s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human BMP2 expression vector containing a Myc-His tag and (B) HEK-293T transfected with empty vector containing a Myc-His tag tissue labeling BMP2 with ab325708 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human BMP2 expression vector containing a Myc-His tag, no staining on (B) HEK-293T 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.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-BMP2 antibody [EPR30246-26] (AB325708)

Indirect ELISA analysis of ab325708 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 : Human BMP2;Human BMP4;Human BMP7.

Antigen concentration : 1000 ng/ml