Cellular metabolism · target hub

DHFR

DHFR (DHFR)

DHFR is a OXPHOS / pentose phosphate on the cellular metabolism axis. Validate function and tissue distribution with UniProt and primary literature. It is indexed under "PPP & one-carbon / folate (representative)" on our pathway page for antibodies, inhibitors, and assay guidance.

Back to pathway overviewSearch: DHFRUniProt (Q9Y243)

Research notes

  • Combine with other genes in the same hub block via genetics or pharmacology to test non-redundant roles of DHFR.
  • Pair metabolic flux with 13C tracing, Seahorse OCR/ECAR, or metabolomics; nutrient withdrawal/refeed controls.
  • Verify antibody clone, compartment, and stimulation; include KD/KO or inhibitor controls.

On the pathway hub (sections)

Genetic lesions & expression context (quick reference)

Type / exampleDomain / context (brief)
Expression contextTissue/cell-type–dependent expression
Rare variantsSporadic variants—require functional validation
Pathway couplingCo-occurs with neighboring nodes on the hub

Naming and prevalence vary by cohort and assay—annotate clinically with COSMIC, ClinVar, OncoKB, and datasheets; research context only.

Assay readouts for DHFR

  • OXPHOS/PPP: DHFR with OCR, ATP, or NADPH readouts.
  • Oligomycin/antimycin A/rotenone pharmacology.
  • Oxidative stress and GSH/GSSG.
  • One-carbon/folate LC-MS for PPP nodes.

DHFR experimental notes

Examples below reflect common literature and public resources (e.g., CCLE, DepMap)—validate genotypes, expression, and passage in your own stocks before committing assays.

[1] Cell lines commonly used for DHFR studies (examples)

  • Tool lines: HeLa, HepG2, Jurkat, HEK293T, A549—screen by DHFR expression and pathway context (CCLE/DepMap).
  • Combine with neighbors (G6PD, PGD, TALDO1, TKT) via KD/pharmacology to test non-redundancy.
  • Overexpression/rescue in HEK293T supports mechanism and IP workflows.
  • Isogenic/CRISPR models separate pathway dependency from bypass survival.

[2] Cell samples for DHFR Western blot

  • Whole-cell lysates—optimize RIPA/NP-40 per antibody; phosphatase inhibitors for phospho blots.
  • Stimulation: serum starvation ± insulin/EGF or amino-acid withdrawal/refeed as relevant.
  • Controls: siRNA/shRNA, CRISPR KO, or inhibitors to validate band specificity.
  • Loading: BCA normalization; subcellular fractionation when needed.

[3] Tissue samples for DHFR Western blot

  • Matched tumor/adjacent frozen tissues after pathology review.
  • Mouse GEMM or xenografts—mind species antibody cross-reactivity.
  • Primary cells/PDCs when ethically approved.
  • FFPE needs specialized extraction; frozen tissue preferred for phospho work.

[4] Cell samples for DHFR immunoprecipitation

  • Tagged overexpression: HEK293T FLAG/HA-DHFR for complex capture.
  • Endogenous IP: high-expression lines; often ≥1–5×10⁶ cells per IP.
  • Stimulation: ligand or nutrient treatments enrich interactions—pilot time courses.
  • Controls: isotype IgG, empty vector, KD/KO negatives.

Human tissues and primary cells require ethics/IRB approval; tumors are heterogeneous—record histotype, site, and preservation conditions.

Bypass & related pathways

Models & genetics note

Overexpression vs endogenous DHFR can differ in dosage, splicing, and compartmentation—in organoids/PDX, record passage, matrix, and drug history.

Quick search presets

DHFRDHFR antibodyDHFR ligand/agonistmetabolism enzyme/transporter antibody

DHFR-related antibodies (keyword-biased)

Adds DHFR keyword bias atop the 细胞代谢 antibody pool—if sparse, use presets above or global search.

Neighboring targets (same hub)

G6PDPGDTALDO1TKTRPIARPEMTHFD1MTHFD2

External databases & resources

FAQ

This content supports research reagents and pathway education—not medical advice. Annotate mutations, drug indications, and protocols with authoritative databases, datasheets, and institutional oversight.

Last reviewed: 2026-05-18

Related on this site

See the cellular metabolism hub for neighboring targets—cross-check PI3K, MAPK, and transmembrane transport pages as needed.

Cellular metabolism·PI3K/Akt/mTOR·MAPK/ERK·Receptor tyrosine kinase·Transmembrane transport