Anti-Aggrecan antibody [RM2036]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Aggrecan with ab315486 at 1/4000 (0.132 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Human cartilage tissue labeling Aggrecan with ab315486 at 1/4000 (0.132 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cartilage. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a ACAN expression vector containing a his tag. (B) HEK-293T transfected with empty vector containing a his tag. tissue labeling Aggrecan with ab315486 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a ACAN expression vector containing a his tag, no staining on (B) HEK-293T transfected with empty vector containing a his tag. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Mouse cartilage tissue labeling Aggrecan with ab315486 at 1/4000 (0.132 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cartilage. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Rat cartilage tissue labeling Aggrecan with ab315486 at 1/10000 (0.053 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cartilage. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Aggrecan with ab315486 at 1/4000 (0.132 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver. The section was incubated with ab315486 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-Aggrecan antibody [RM2036] (AB315486)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Aggrecan with ab315486 at 1/10000 (0.053 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver. The section was incubated with ab315486 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

• IP

Supplier Data

Immunoprecipitation - Anti-Aggrecan antibody [RM2036] (AB315486)

Aggrecan was immunoprecipitated from 0.35 mg Mouse articular cartilage tissue lysate with ab315486 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315486 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Aggrecan antibody [RM2036] (ab315486) at 1/1000 dilution

Lane 1:

Mouse articular cartilage tissue lysate

Lane 2:

ab315486 at 1/30 IP in Mouse articular cartilage tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315486 in Mouse articular cartilage tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Aggrecan antibody [RM2036] (AB315486)

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

Negative control : liver.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11425904, 16188468).

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

All lanes:

Western blot - Anti-Aggrecan antibody [RM2036] (ab315486) at 1/1000 dilution

Lane 1:

Human synovial fluid lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50-350 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Aggrecan antibody [RM2036] (AB315486)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11425904, 16188468).

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-Aggrecan antibody [RM2036] (ab315486) at 1/1000 dilution

All lanes:

Human articar cartilage tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50-350 kDa,36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Aggrecan antibody [RM2036] (AB315486)

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

Negative control : liver, spleen.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11425904, 16188468).

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

Exposure time : Lanes 1-3 : 1second, lanes 4-5 : 3 seconds.

All lanes:

Western blot - Anti-Aggrecan antibody [RM2036] (ab315486) at 1/1000 dilution

Lane 1:

Mouse articar cartilage tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Rat articar cartilage tissue lysate at 20 µg

Lane 5:

Rat spleen tissue 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: 50-350 kDa

false

• WB

Supplier Data

Western blot - Anti-Aggrecan antibody [RM2036] (AB315486)

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

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Aggrecan antibody [RM2036] (ab315486) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human ACAN expression vector containing a His-tag, 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: 75 kDa,36 kDa

false

Exposure time: 1s