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Glycerol Kinase Recombinant Rabbit Monoclonal Antibody [JE55-12]

Usd: 385

Specification

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Catalog# ET7110-96

Glycerol Kinase Recombinant Rabbit Monoclonal Antibody [JE55-12]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Glycerol Kinase Recombinant Rabbit Monoclonal Antibody [JE55-12]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Glycerol kinase aa 440-559 / 559.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Target Molecular Weight

Predicted band size 61/57 kDa

Positive Control

A549, rat kidney tissue lysate, rat testis tissue, human liver tissue, human kidney tissue, mouse testis tissue, 293.

Conjugation

unconjugated

Clone Number

JE55-12

RRID

AB_3071055

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

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

Glycerol kinase, encoded by the gene GK, is a phosphotransferase enzyme involved in triglycerides and glycerophospholipids synthesis. Glycerol kinase catalyzes the transfer of a phosphate from ATP to glycerol thus forming glycerol 3-phosphate. Adipocytes lack glycerol kinase so they cannot metabolize the glycerol produced during triacyl glycerol degradation. This glycerol is instead shuttled to the liver via the blood where it is: Phosphorylated by glycerol kinase to glycerol 3-phosphate. Converted from glycerol 3-phosphate to dihydroxyacetone phosphate (DHAP) via glycerol 3-phosphate dehydrogenase. DHAP can participate in glycolysis or gluconeogenesis. This protein may use the morpheein model of allosteric regulation. Glycerol Kinase (alternative name, ATP:glycerol 3-phosphotransferase or Glycerokinase) adopts a ribonuclease H-like fold consisting of an alpha-beta 2-layer sandwich of CATH family 3.30.420.40. As of March 2010, there were 20 structures of this protein in the PDB, most of which are homodimeric.

Background References

1. Arrieta F. et. al. Glycerol kinase deficiency in adult patient: hypertriglyceridemia resistance to diet and pharmacological treatment. Nutr Hosp. 2018 Aug.

2. Bellerose MM. et. al. Common Variants in the Glycerol Kinase Gene Reduce Tuberculosis Drug Efficacy. mBio. 2019 Jul.

Sequence Similarity

Belongs to the FGGY kinase family.

Tissue Specificity

Highly expressed in the liver, kidney and testis. Isoform 2 and isoform 3 are expressed specifically in testis and fetal liver, but not in the adult liver.

Subcellular Location

Mitochondrion outer membrane, cytoplasm.

UNIPROT

SwissProt: P32189 Human

SwissProt: Q64516 Mouse

SwissProt: Q63060 Rat

Synonyms

ATP glycerol 3 phosphotransferase antibody

ATP:glycerol 3 phosphotransferase antibody

ATP:glycerol 3-phosphotransferase antibody

D930012N15Rik antibody

GK antibody

GK1 antibody

GKD antibody

GLPK_HUMAN antibody

Glycerokinase antibody

Glycerol kinase antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Glycerol Kinase on different lysates with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si Glycerol Kinase cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 61/57 kDa<br />Observed band size: 61/57 kDa<br /><br />Exposure time: 1 minute; ECL: K1802;<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/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</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.

    ☑ Knockdown (KD)

    Western blot analysis of Glycerol Kinase on different lysates with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/1,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si Glycerol Kinase cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 61/57 kDa
    Observed band size: 61/57 kDa

    Exposure time: 1 minute; ECL: K1802;

    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 (ET7110-96) 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.

  • Western blot analysis of Glycerol Kinase on different lysates with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/1,000 dilution.<br /><br />Lane 1: rat kidney tissue lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 6 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>, 1/1,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 61.2 kDa<br />Observed band size: 61/57 kDa

    Western blot analysis of Glycerol Kinase on different lysates with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/1,000 dilution.

    Lane 1: rat kidney tissue lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 6 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET7110-96, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 61.2 kDa
    Observed band size: 61/57 kDa

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-96) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-96) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-96) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Glycerol Kinase antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Glycerol Kinase antibody (ET7110-96) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer(pH 8.0-8.4))for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-96) at 1/200 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Glycerol Kinase was done on 293 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7110-96" style="font-weight: bold;text-decoration: underline;">ET7110-96</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Glycerol Kinase was done on 293 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-96, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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