Anti-MTHFD2 antibody (ab56772)
Key features and details
- Mouse monoclonal to MTHFD2
- Suitable for: WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-MTHFD2 antibody
See all MTHFD2 primary antibodies -
Description
Mouse monoclonal to MTHFD2 -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-P HumanWB Human
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Immunogen
Recombinant full length protein, corresponding to amino acids 16-345 of Human MTHFD2
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Positive control
- WB: HEK293T, HepG2 and HeLa cell lysates. IHC-P: Human lymphoma tissue. Flow Cyt: HepG2 cells.
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General notes
This product was changed from ascites to tissue culture supernatant on 24/1/19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.4 -
Concentration information loading... -
Purity
Tissue culture supernatant -
Purification notes
Purified from TCS. -
Clonality
Monoclonal -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab56772 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| Flow Cyt |
Human
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| IHC-P |
Human
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| WB |
Human
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| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 37 kDa.
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| IHC-P |
Use at an assay dependent concentration.
|
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| Flow Cyt |
Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 37 kDa. |
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IHC-P
Use at an assay dependent concentration. |
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Flow Cyt
Use at an assay dependent concentration. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
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Target
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Sequence similarities
Belongs to the tetrahydrofolate dehydrogenase/cyclohydrolase family. -
Developmental stage
Expressed only in developing normal tissues. -
Cellular localization
Mitochondrion. - Information by UniProt
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Database links
- Entrez Gene: 10797 Human
- Entrez Gene: 441024 Human
- Omim: 604887 Human
- SwissProt: P13995 Human
- SwissProt: Q9H903 Human
- Unigene: 469030 Human
- Unigene: 721011 Human
- Unigene: 740700 Human
see all -
Alternative names
- Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial antibody
- Descriptions antibody
- Methenyltetrahydrofolate cyclohydrolase antibody
see all
Images
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Western blot - Anti-MTHFD2 antibody (ab56772)All lanes : Anti-MTHFD2 antibody (ab56772) at 1/500 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : MTHFD2 knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab56772 observed at 36 kDa. Red - loading control ab52901 observed at kDa.
ab56772 Anti-MTHFD2 antibody was shown to specifically react with MTHFD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266383 (knockout cell lysate ab258528) was used. Wild-type and MTHFD2 knockout samples were subjected to SDS-PAGE. ab56772 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD2 antibody (ab56772)MTHFD2 antibody (ab56772) used in immunohistochemistry at 3μg/ml on formalin fixed and paraffin embedded human lymphoma tissue.
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Flow Cytometry - Anti-MTHFD2 antibody (ab56772)Overlay histogram showing HepG2 cells stained with ab56772 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56772, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using the ascites version of the product. -
Western blot - Anti-MTHFD2 antibody (ab56772)MTHFD2 antibody (ab56772) at 1μg/lane + HepG2 cell lysate at 25μg/lane.
This image was generated using the ascites version of the product.
Protocols
Datasheets and documents
References (6)
ab56772 has been referenced in 6 publications.
- Nishimura T et al. Cancer stem-like properties and gefitinib resistance are dependent on purine synthetic metabolism mediated by the mitochondrial enzyme MTHFD2. Oncogene 38:2464-2481 (2019). PubMed: 30532069
- Green NH et al. MTHFD2 links RNA methylation to metabolic reprogramming in renal cell carcinoma. Oncogene 38:6211-6225 (2019). PubMed: 31289360
- Gustafsson R et al. Crystal Structure of the Emerging Cancer Target MTHFD2 in Complex with a Substrate-Based Inhibitor. Cancer Res 77:937-948 (2017). PubMed: 27899380
- Pikman Y et al. Targeting MTHFD2 in acute myeloid leukemia. J Exp Med 213:1285-306 (2016). PubMed: 27325891
- Gustafsson Sheppard N et al. The folate-coupled enzyme MTHFD2 is a nuclear protein and promotes cell proliferation. Sci Rep 5:15029 (2015). WB . PubMed: 26461067
- Nilsson R et al. Metabolic enzyme expression highlights a key role for MTHFD2 and the mitochondrial folate pathway in cancer. Nat Commun 5:3128 (2014). WB ; Human . PubMed: 24451681
