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AKR1C1 / AKR1C2 Recombinant Rabbit Monoclonal Antibody [JE33-84]

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Specification

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Catalog# HA722694

AKR1C1 / AKR1C2 Recombinant Rabbit Monoclonal Antibody [JE33-84]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

AKR1C1 / AKR1C2 Recombinant Rabbit Monoclonal Antibody [JE33-84]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human AKR1C1 aa 51-100 / 323.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 37 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, Human liver tissue lysate, Mouse liver tissue lysate, Mouse testis tissue lysate, HepG2.

Conjugation

unconjugated

Clone Number

JE33-84

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Aldo-keto reductase family 1 (AKR1) is a family of aldo-keto reductase enzymes that is involved in steroid metabolism. It includes the AKR1C and AKR1D subgroups, which respectively consist of AKR1C1–AKR1C4 and AKR1D1. Together with short-chain dehydrogenase/reductases (SDRs), these enzymes catalyze oxidoreductions, act on the C3, C5, C11, C17 and C20 positions of steroids, and function as 3α-HSDTooltip 3α-Hydroxysteroid dehydrogenases, 3β-HSDsTooltip 3β-Hydroxysteroid dehydrogenases, 5β-reductases, 11β-HSDsTooltip 11β-Hydroxysteroid dehydrogenases, 17β-HSDsTooltip 17β-hydroxysteroid dehydrogenases, and 20α-HSDsTooltip 20α-Hydroxysteroid dehydrogenases, respectively. The AKR1C enzymes act as 3-, 17- and 20-ketosteroid reductases, while AKR1D1 acts as the sole 5β-reductase in humans.

Background References

1. Zuo X et al. AKR1C1 Protects Corneal Epithelial Cells Against Oxidative Stress-Mediated Ferroptosis in Dry Eye. Invest Ophthalmol Vis Sci. 2022 Sep

2. Kaftalli J et al. AKR1C1 and hormone metabolism in lipedema pathogenesis: a computational biology approach. Eur Rev Med Pharmacol Sci. 2023 Dec

Subcellular Location

Cytoplasm, cytosol.

UNIPROT

SwissProt: Q04828 Human

SwissProt: P52895 Human

Synonyms

20 alpha (3 alpha) hydroxysteroid dehydrogenase antibody

20-alpha-HSD antibody

20-alpha-hydroxysteroid dehydrogenase antibody

3-alpha-HSD3 antibody

AKR1C1 antibody

AKR1C2 antibody

Aldo-keto reductase family 1 member C1 antibody

Aldo-keto reductase family 1 member C2 antibody

Chlordecone reductase homolog HAKRC antibody

Chlordecone reductase homolog HAKRD antibody

Expand

Images

  • Western blot analysis of AKR1C1 / AKR1C2 on different lysates with Rabbit anti-AKR1C1 / AKR1C2 antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: HepG2 cell lysate (10 µg/Lane)<br />Lane 3: Human liver tissue lysate (20 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 37 kDa<br />Observed band size: 37 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of AKR1C1 / AKR1C2 on different lysates with Rabbit anti-AKR1C1 / AKR1C2 antibody (HA722694) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: HepG2 cell lysate (10 µg/Lane)
    Lane 3: Human liver tissue lysate (20 µg/Lane)
    Lane 4: Mouse liver tissue lysate (20 µg/Lane)

    Predicted band size: 37 kDa
    Observed band size: 37 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722694) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of AKR1C1 / AKR1C2 on different lysates with Rabbit anti-AKR1C1 / AKR1C2 antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: Mouse testis tissue lysate (30 µg/Lane)<br />Lane 3: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 4: 293T cell lysate (negative) (20 µg/Lane)<br /><br />Predicted band size: 37 kDa<br />Observed band size: 37 kDa<br /><br />Exposure time: 12 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of AKR1C1 / AKR1C2 on different lysates with Rabbit anti-AKR1C1 / AKR1C2 antibody (HA722694) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: Mouse testis tissue lysate (30 µg/Lane)
    Lane 3: Mouse liver tissue lysate (30 µg/Lane)
    Lane 4: 293T cell lysate (negative) (20 µg/Lane)

    Predicted band size: 37 kDa
    Observed band size: 37 kDa

    Exposure time: 12 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722694) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling AKR1C1 / AKR1C2 with Rabbit anti-AKR1C1 / AKR1C2 antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKR1C1 / AKR1C2 antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling AKR1C1 / AKR1C2 with Rabbit anti-AKR1C1 / AKR1C2 antibody (HA722694) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKR1C1 / AKR1C2 antibody (HA722694) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HepG2 cells labeling AKR1C1 / AKR1C2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722694" style="font-weight: bold;text-decoration: underline;">HA722694</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling AKR1C1 / AKR1C2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722694, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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