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IDH2 Recombinant Rabbit Monoclonal Antibody [JA55-31]

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Specification

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Catalog# ET1704-93

IDH2 Recombinant Rabbit Monoclonal Antibody [JA55-31]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

IDH2 Recombinant Rabbit Monoclonal Antibody [JA55-31]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human IDH2 aa 403-452 / 452.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 51 kDa

Positive Control

THP-1 cell lysate, Jurkat cell lysate, MCF7 cell lysate, C2C12 cell lysate, L6 cell lysate, human liver tissue, human tonsil tissue, human prostate tissue, rat heart tissue, mouse heart tissue.

Conjugation

unconjugated

Clone Number

JA55-31

RRID

AB_3070535

Product Features

Form

Liquid

Concentration

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

  • 1:2,000

  • IHC-P

  • 1:200

  • IF-Cell

  • 1:100

Target

Function

Isocitrate dehydrogenase [NADP], mitochondrial is an enzyme that in humans is encoded by the IDH2 gene. Isocitrate dehydrogenases are enzymes that catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer. The protein encoded by the IDH2 gene is the NADP(+)-dependent isocitrate dehydrogenase found in the mitochondria. It plays a role in intermediary metabolism and energy production. This protein may tightly associate or interact with the pyruvate dehydrogenase complex. Somatic mosaic mutations of this gene have also been found associated to Ollier disease and Maffucci syndrome.

Background References

1. Cerchione C et al. IDH1/IDH2 Inhibition in Acute Myeloid Leukemia. Front Oncol. 2021 Mar

2. Issa GC et al. Acute myeloid leukemia with IDH1 and IDH2 mutations: 2021 treatment algorithm. Blood Cancer J. 2021 Jun

Sequence Similarity

Belongs to the isocitrate and isopropylmalate dehydrogenases family.

Post-translational Modification

Acetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3.

Subcellular Location

Mitochondrion.

UNIPROT

SwissProt: P48735 Human

SwissProt: P54071 Mouse

SwissProt: P56574 Rat

Synonyms

D2HGA2 antibody

ICD-M antibody

IDH antibody

IDH2 antibody

IDHM antibody

IDHP_HUMAN antibody

IDP antibody

IDPM antibody

Isocitrate dehydrogenase [NADP], mitochondrial antibody

Isocitrate dehydrogenase 2 (NADP+), mitochondrial antibody

Expand

Images

  • Western blot analysis of IDH2 on different lysates with Rabbit anti-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/2,000 dilution.<br /><br />Lane 1: THP-1 cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: C2C12 cell lysate<br />Lane 5: L6 cell lysate<br /><br />Predicted band size: 51 kDa<br />Observed band size: 45 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/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/2,000 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 IDH2 on different lysates with Rabbit anti-IDH2 antibody (ET1704-93) at 1/2,000 dilution.

    Lane 1: THP-1 cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: C2C12 cell lysate
    Lane 5: L6 cell lysate

    Predicted band size: 51 kDa
    Observed band size: 45 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 (ET1704-93) at 1/2,000 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of IDH2 on different lysates with Rabbit anti-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-IDH2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 1 minute; 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/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 IDH2 on different lysates with Rabbit anti-IDH2 antibody (ET1704-93) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-IDH2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 51 kDa
    Observed band size: 45 kDa

    Exposure time: 1 minute; 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 (ET1704-93) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/200 dilution for 1 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-IDH2 antibody (ET1704-93) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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 (ET1704-93) at 1/200 dilution for 1 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 heart tissue with Rabbit anti-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/200 dilution for 1 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 heart tissue with Rabbit anti-IDH2 antibody (ET1704-93) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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 (ET1704-93) at 1/200 dilution for 1 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.

  • Immunocytochemistry analysis of THP-1 cells labeling IDH2 with Rabbit anti-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IDH2 antibody (<a href="/products/ET1704-93" style="font-weight: bold;text-decoration: underline;">ET1704-93</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 THP-1 cells labeling IDH2 with Rabbit anti-IDH2 antibody (ET1704-93) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IDH2 antibody (ET1704-93) 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 (HA601187 , 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.

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Citation