Anti-Clusterin antibody [RM1292]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Clusterin with ab323182 at 1/1000 (0.496 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : almost no staining on human liver.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Clusterin with ab323182 at 1/1000 (0.496 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Clusterin with ab323182 at 1/1000 (0.496 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Clusterin was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab323182 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323182 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab323182 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323182 in HeLa whole cell 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: 8s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Mouse adrenal gland tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on medulla of mouse adrenal gland.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Rat adrenal gland tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on medulla of rat adrenal gland.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : almost no staining on rat skeletal muscle.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Clusterin with ab323182 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on the progenitor cells of rat colon crypt.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Clusterin with ab323182 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : almost no staining on mouse skeletal muscle.
The section was incubated with ab323182 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-Clusterin antibody [RM1292] (AB323182)

Clusterin was immunoprecipitated from 0.35 mg Rat plasma lysate with ab323182 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323182 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

Rat plasma lysate at 10 µg

Lane 2:

ab323182 at 1/30 IP in rat plasma lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323182 in rat plasma 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: 67s

• IP

Supplier Data

Immunoprecipitation - Anti-Clusterin antibody [RM1292] (AB323182)

Clusterin was immunoprecipitated from 0.35 mg Mouse plasma lysate with ab323182 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323182 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

Mouse plasma lysate at 10 µg

Lane 2:

ab323182 at 1/30 IP in mouse plasma lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323182 in mouse plasma 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: 50s

• WB

Supplier Data

Western blot - Anti-Clusterin antibody [RM1292] (AB323182)

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

Low expression tissue : skeletal muscle

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 8373947).

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

All lanes:

Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Lane 3:

Human skeletal muscle 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: 62 kDa,38 kDa,36 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Clusterin antibody [RM1292] (AB323182)

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

Negative control : Jurkat

In Western blot, ab323182 was shown to bind specifically to Clusterin. Target of interest was observed at 62 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Clusterin knockout cell line (lane 2).

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 8373947).

Recombinant multiclonal ab323182 performs better than polyclonal in WB application.

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

Recombinant multiclonal ab323182 : 114 seconds; Polyclonal ab69644 : 180 seconds

All lanes:

Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

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

Lane 2:

Clusterin knockout HeLa whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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: 62 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Clusterin antibody [RM1292] (AB323182)

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

Low expression tissue : skeletal muscle

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 8373947).

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

Exposure time : Lane 1, 2 : 15 seconds; Lane 3, 4 : 37 seconds

All lanes:

Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat skeletal muscle 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: 62 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Clusterin antibody [RM1292] (AB323182)

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

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 8373947).

All lanes:

Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

Lane 1:

Mouse plasma lysate at 20 µg

Lane 2:

Rat plasma 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: 38 kDa

false

Exposure time: 48s