Anti-AKR1C3 antibody [EPR28747-41]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized TF-1 (human erythroleukemia erythroblast) cells labelling AKR1C3 with ab316864 at 1/50 (10.0 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear and cytoplasmic staining in TF-1 cell line.
Low expression : HCT 116 (PMID : 21215737).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human cardiac muscle (PMID : 33352741).
The section was incubated with ab316864 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon (PMID : 33352741).
The section was incubated with ab316864 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hepatocellular carcinoma.
The section was incubated with ab316864 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell, Left) / TF-1 (human erythroleukemia erythroblast, Right) cells labelling AKR1C3 with ab316864 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : HCT 116.

• IP

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Immunoprecipitation - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

AKR1C3 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate with ab316864 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316864 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : ab316864 IP in A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316864 in A549 whole cell lysate

All lanes:

Immunoprecipitation - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/30 dilution

All lanes:

A549 (human lung carcinoma 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: 6s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon.
The section was incubated with ab316864 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-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver.
The section was incubated with ab316864 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-AKR1C3 antibody [EPR28747-41] (AB316864)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cardiac muscle.
The section was incubated with ab316864 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

Supplier Data

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Low expression : HCT 116 (PMID : 21215737, PMID : 35388063)

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

Lane 1:

TF-1 (human Erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 2:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellar carcinoma 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: 36 kDa,124 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Low expression : muscle (PMID : 18635746)

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

All lanes:

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

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 1/100000 dilution

Observed band size: 36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Low expression : muscle (PMID : 18635746)

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

All lanes:

Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse 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 1/100000 dilution

Observed band size: 36 kDa

false

Exposure time: 10s

• Dot

Lab

Dot Blot - Anti-AKR1C3 antibody [EPR28747-41] (AB316864)

Dot blot analysis of AKR1C3 using ab316864 (Lane 1-4) at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 Dilution] dilution and Dot blot analysis of AKR1C3 using ab213204 (lane 5-8) at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution.

Lane 1&5 : His tagged AKR1C1 recombinant protein
Lane 2&6 : His tagged AKR1C2 recombinant protein
Lane 3&7 : AKR1C3 recombinant protein without tag
Lane 4&8 : His tagged AKR1C4 recombinant protein

Lanes 1 - 4:

Dot Blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

Lanes 5 - 8:

Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/en-us/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>) at 1/1000 dilution

Lanes 1 and 5:

His tagged AKR1C1 recombinant protein

Lanes 2 and 6:

His tagged AKR1C2 recombinant protein

Lanes 3 and 7:

AKR1C3 recombinant protein without tag

Lanes 4 and 8:

His tagged AKR1C4 recombinant protein

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>)

false

Exposure time: 180s