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MTHFD2 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER2001-62

MTHFD2 Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

MTHFD2 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within human MTHFD2 aa 300-350.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 38 kDa

Positive Control

293 cell lysates, HEK-293, human breast cancer tissue, human tonsil tissue.

Conjugation

unconjugated

RRID

AB_3069539

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), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:500

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:200-1:500

  • FC

  • 1:500

Target

Function

Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial is an enzyme that in humans is encoded by the MTHFD2 gene. This gene encodes a nuclear-encoded mitochondrial bifunctional enzyme with methylenetetrahydrofolate dehydrogenase and methenyltetrahydrofolate cyclohydrolase activities. The enzyme functions as a homodimer and is unique in its absolute requirement for magnesium and inorganic phosphate. Formation of the enzyme-magnesium complex allows binding of NAD. Alternative splicing results in two different transcripts, one protein-coding and the other not protein-coding. This gene has a pseudogene on chromosome 7.

Background References

1. Green NH. et. al. MTHFD2 links RNA methylation to metabolic reprogramming in renal cell carcinoma. Oncogene. 2019 Aug.

Subcellular Location

Mitochondrion.

UNIPROT

SwissProt: P13995 Human

Synonyms

Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial antibody

Descriptions antibody

Methenyltetrahydrofolate cyclohydrolase antibody

methylene tetrahydrofolate dehydrogenase (NAD+ dependent) antibody

methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2 antibody

methylenetetrahydrofolate dehydrogenase (NADP+ dependent) antibody

methylenetetrahydrofolate dehydrogenase 2 antibody

MGC82516 antibody

MTDC_HUMAN antibody

MTHFD2 antibody

Expand

Images

  • Western blot analysis of MTHFD2 on 293 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MTHFD2 on 293 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER2001-62, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HEK-293 cells labeling MTHFD2 with Rabbit anti-MTHFD2 antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>) at 1/50 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-MTHFD2 antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>) at 1/50 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 HEK-293 cells labeling MTHFD2 with Rabbit anti-MTHFD2 antibody (ER2001-62) at 1/50 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-MTHFD2 antibody (ER2001-62) at 1/50 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.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-MTHFD2 antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>) at 1/500 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/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>) at 1/500 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 breast cancer tissue with Rabbit anti-MTHFD2 antibody (ER2001-62) at 1/500 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 (ER2001-62) at 1/500 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 tonsil tissue with Rabbit anti-MTHFD2 antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</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/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</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 tonsil tissue with Rabbit anti-MTHFD2 antibody (ER2001-62) 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 (ER2001-62) 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.

  • Flow cytometric analysis of HEK-293 cells labeling MTHFD2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ER2001-62" style="font-weight: bold;text-decoration: underline;">ER2001-62</a>, 1/500) (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 HEK-293 cells labeling MTHFD2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ER2001-62, 1/500) (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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Application: WB,IF-Cell,IHC-P,FC,IP

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated