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IDH1 Recombinant Rabbit Monoclonal Antibody [JE34-27]

Usd: 385

Specification

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Catalog# HA722696

IDH1 Recombinant Rabbit Monoclonal Antibody [JE34-27]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

IDH1 Recombinant Rabbit Monoclonal Antibody [JE34-27]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human IDH1 aa 51-100 / 414.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 47 kDa

Positive Control

HeLa cell lysate, Raji cell lysate, HepG2 cell lysate, Neuro-2a cell lysate, C6 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, human prostate cancer tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

JE34-27

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:1,000-1:2,000

  • IHC-P

  • 1:3,000

  • IP

  • 1-2μg/sample

Target

Function

Isocitrate dehydrogenase 1 (NADP+), soluble is an enzyme that in humans is encoded by the IDH1 gene on chromosome 2. Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which uses 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 this gene is the NADP+-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence. The presence of this enzyme in peroxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2,4-dienoyl-CoAs to 3-enoyl-CoAs, as well as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production. Alternatively spliced transcript variants encoding the same protein have been found for this gene.

Background References

1. Tian W et al. Recent advances of IDH1 mutant inhibitor in cancer therapy. Front Pharmacol. 2022 Aug

2. Platten M et al. A vaccine targeting mutant IDH1 in newly diagnosed glioma. Nature. 2021 Apr

Subcellular Location

Cytoplasm, cytosol, Peroxisome.

UNIPROT

SwissProt: O75874 Human

SwissProt: O88844 Mouse

SwissProt: P41562 Rat

Synonyms

Cytosolic NADP isocitrate dehydrogenase antibody

Cytosolic NADP-isocitrate dehydrogenase antibody

Epididymis luminal protein 216 antibody

Epididymis secretory protein Li 26 antibody

HEL-216 antibody

HEL-S-26 antibody

ICDH antibody

IDCD antibody

IDH antibody

IDH1 antibody

Expand

Images

  • Western blot analysis of IDH1 on different lysates with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Raji cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: Neuro-2a cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: Mouse kidney tissue lysate<br />Lane 7: Rat kidney tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 47 kDa<br />Observed band size: 47 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</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 IDH1 on different lysates with Rabbit anti-IDH1 antibody (HA722696) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Raji cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: Neuro-2a cell lysate
    Lane 5: C6 cell lysate
    Lane 6: Mouse kidney tissue lysate
    Lane 7: Rat kidney tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 47 kDa
    Observed band size: 47 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 (HA722696) 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of IDH1 on different lysates with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-IDH1 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 47 kDa<br />Observed band size: 47 kDa<br /><br />Exposure time: 6 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/2,000 dilution was used in K1803 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 IDH1 on different lysates with Rabbit anti-IDH1 antibody (HA722696) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-IDH1 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

    Exposure time: 6 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 (HA722696) at 1/2,000 dilution was used in 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 prostate cancer tissue with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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 prostate cancer tissue with Rabbit anti-IDH1 antibody (HA722696) at 1/3,000 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 (HA722696) at 1/3,000 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 kidney tissue with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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 kidney tissue with Rabbit anti-IDH1 antibody (HA722696) at 1/3,000 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 (HA722696) at 1/3,000 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 kidney tissue with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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 kidney tissue with Rabbit anti-IDH1 antibody (HA722696) at 1/3,000 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 (HA722696) at 1/3,000 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 rat kidney tissue with Rabbit anti-IDH1 antibody (<a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a>) at 1/3,000 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 rat kidney tissue with Rabbit anti-IDH1 antibody (HA722696) at 1/3,000 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 (HA722696) at 1/3,000 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.

  • IDH1 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722696" style="font-weight: bold;text-decoration: underline;">HA722696</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 8 seconds; ECL: K1802

    IDH1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722696 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722696 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA722696 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA722696 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 8 seconds; ECL: K1802

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