Anti-Deptor antibody [EPR26159-220]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Deptor antibody [EPR26159-220] (AB309531)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Deptor with ab309531 at 1/200 (2.56 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Low expression tissue : Nearly no staining on human kidney.The section was incubated with ab309531 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) was used. 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-Deptor antibody [EPR26159-220] (AB309531)

Immunohistochemical analysis of paraffin-embedded Human prostatic hype tissue labeling Deptor with ab309531 at 1/200 (2.56 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human prostatic hyperplasia (PMID : 31685947).The section was incubated with ab309531 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Deptor antibody [EPR26159-220] (AB309531)

Immunohistochemical analysis of paraffin-embedded Human thyroid tissue labeling Deptor with ab309531 at 1/200 (2.56 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human thyroid.The section was incubated with ab309531 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) was used. 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-Deptor antibody [EPR26159-220] (AB309531)

Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling Deptor with ab309531 at 1/200 (2.56 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human endometrium.The section was incubated with ab309531 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Deptor antibody [EPR26159-220] (AB309531)

Deptor was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab309531 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309531 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab309531 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309531 in MCF7 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 58 seconds

All lanes:

Immunoprecipitation - Anti-Deptor antibody [EPR26159-220] (ab309531) at 1/30 dilution

All lanes:

MCF7 (human breast adenocarcinoma epithelial cell) 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: 58s

• WB

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Western blot - Anti-Deptor antibody [EPR26159-220] (AB309531)

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

Low expression : PC-3 (PMID : 19446321)

The identity of the lower MW band at approximately 35kDa is unknown. The band may represent a Deptor degradation product (PMID : 34519269).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Deptor antibody [EPR26159-220] (ab309531) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma 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 1/100000 dilution

Observed band size: 48 kDa,34 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Deptor antibody [EPR26159-220] (AB309531)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the lower MW band at approximately 35kDa is unknown. The band may represent a Deptor degradation product (PMID : 34519269).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Deptor antibody [EPR26159-220] (ab309531) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa serum starved overnight, 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: 48 kDa,34 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Deptor antibody [EPR26159-220] (AB309531)

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

The identity of the lower MW band at approximately 35kDa is unknown. The band may represent a Deptor degradation product (PMID : 34519269).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Exposure time :

Lane 1-3 : 3 minutes

Lane 4 : 81 seconds

All lanes:

Western blot - Anti-Deptor antibody [EPR26159-220] (ab309531) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

AR42J (rat pancreatic tumor 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 1/100000 dilution

Observed band size: 48 kDa,34 kDa

true