MTHFD2 Recombinant Rabbit Monoclonal Antibody [PSH24-73]
Catalog# HA724469
MTHFD2 Recombinant Rabbit Monoclonal Antibody [PSH24-73]
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WB
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IF-Cell
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IHC-P
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
MTHFD2 Recombinant Rabbit Monoclonal Antibody [PSH24-73]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human MTHFD2 aa 51-100.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 38 kDa
Positive Control
HEK-293 (Human embryonic kidney cell)cell lysate, MDA-MB-231 (Human breast cancer cell)cell lysate, HeLa (Human cervical adenocarcinoma cell)cell lysate, 786-0 (Human renal clear cell adenocarcinoma cell)cell lysate, MEF (Mouse embryonic fibroblast)cell lysate, Mouse spleen tissue lysate, Mouse testis tissue lysate, Rat spleen tissue lysate, Rat testis tissue lysate.
Conjugation
unconjugated
Clone Number
PSH24-73
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*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:100
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IHC-P
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1:1,000
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FC
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1:1,000
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IP
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1-2μg/sample
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
2. Hitzel J. et. al. Oxidized phospholipids regulate amino acid metabolism through MTHFD2 to facilitate nucleotide release in endothelial cells. Nat Commun. 2018 Jun
Subcellular Location
Mitochondrion.
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
ExpandImages
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Western blot analysis of MTHFD2 on different lysates with Rabbit anti-MTHFD2 antibody (HA724469) at 1/5,000 dilution.
Lane 1: HEK-293 (Human embryonic kidney cell)cell lysate
Lane 2: MDA-MB-231 (Human breast cancer cell)cell lysate
Lane 3: HeLa (Human cervical adenocarcinoma cell)cell lysate
Lane 4: 786-0 (Human renal clear cell adenocarcinoma cell)cell lysate
Lane 5: MEF (Mouse embryonic fibroblast)cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 8 seconds ; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 38 kDa
Observed band size: 35 kDa -
Western blot analysis of MTHFD2 on different lysates with Rabbit anti-MTHFD2 antibody (HA724469) at 1/5,000 dilution.
Lane 1: Mouse spleen tissue lysate
Lane 2: Mouse testis tissue lysate
Lane 3: Rat spleen tissue lysate
Lane 4: Rat testis tissue lysate
Lysates/proteins at 30 µg/Lane.
Exposure time: 8 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 38 kDa
Observed band size: 35 kDa -
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: HEK-293 (Human embryonic kidney cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724469, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). -
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: HEK-293 (Human embryonic kidney cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724469, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: The mitochondria dyes was Mito-Tracker (Beyotime, Red). The nuclear counterstain was DAPI (Blue). -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Tonsil
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Breast cancer
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Spleen
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Spleen
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Spleen
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724469, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Flow Cytometry (Intra)
Species: Human
Sample: HEK-293 (Human embryonic kidney cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724469(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
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Application: Flow Cytometry (Intra)
Species: Mouse
Sample: MEF (Mouse embryonic fibroblast)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724469(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
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Immunoprecipitation (IP)
MTHFD2 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724469 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724469 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724469 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724469 in HeLa cell lysate
Exposure time: 5 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724469, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 38 kDa
Observed band size: 35 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"