Customize ufm-1 Antibody

Description

What is UFM1 Antibody?

UFM1 antibodies are immunoreagents designed to recognize and bind to UFM1 or its conjugates. These antibodies enable researchers to investigate UFM1's expression, localization, and functional roles in cellular pathways. Two commercially available UFM1 antibodies are widely cited:

Research Applications

UFM1 antibodies are pivotal in studying UFMylation's role in disease mechanisms:

Cancer Research

Gastric Cancer: UFM1 overexpression suppresses tumor invasion and metastasis by modulating migration pathways . Knockdown of UFM1 in gastric cancer cells (e.g., HGC-27) increased invasiveness in vitro and in vivo .
Oral Squamous Cell Carcinoma (OSCC): High UFM1 expression correlates with poor prognosis and immune infiltration (e.g., Th17 and cytotoxic cells) . Inhibition of UFM1 reduced OSCC cell proliferation and migration .

Immune Regulation

PD-L1 Modulation: UFM1 antibodies identified UFL1 (E3 ligase for UFMylation) as a regulator of PD-L1 stability. Loss of UFL1 increases PD-L1 membrane localization, impairing CD8+ T cell-mediated tumor immunity .
Antiviral Response: UFM1 antibodies confirmed UFL1's role in stabilizing STING, enhancing cGAS-STING pathway activity during DNA virus infection .

Specificity and Sensitivity

Cell Signaling #63445: Validated in WB and IP, detecting endogenous UFM1 at ~10 kDa .
Proteintech 15883-1-AP: Recognizes UFM1 in human/mouse tissues (e.g., liver, lung) and cell lines (HEK-293, HepG2) . Bands >100 kDa may indicate UFM1-conjugated proteins .

Immunohistochemistry (IHC)

Proteintech’s antibody demonstrated reactivity in human lung cancer tissues, with optimal antigen retrieval using TE buffer (pH 9.0) .

Clinical and Therapeutic Implications

Biomarker Potential: Elevated UFM1 in OSCC correlates with poor survival and immune evasion .
Therapeutic Target: Small-molecule inhibitors targeting UFSP2 (a UFM1 protease) enhanced PD-L1 degradation, improving anti-PD1 therapy efficacy in preclinical models .

Future Directions

Research priorities include:

Mapping UFM1’s interaction networks in ER-phagy and autophagy .
Developing UFM1-targeted therapies for cancers with dysregulated UFMylation .

Product Specs

Buffer

Preservative: 0.03% ProClin 300; Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 weeks (made-to-order)

Synonyms

ufm-1 antibody; tag-277 antibody; ZK652.3 antibody; Ubiquitin-fold modifier 1 antibody

Target Names

ufm-1

Uniprot No.

P34661

Target Background

Function

UFM1 (Ubiquitin-fold modifier 1) is a ubiquitin-like modifier covalently attached to substrate proteins as a monomer or lysine-linked polymer. This process, termed ufmylation, requires the UFM1-activating enzyme UBA5 (E1), the UFM1-conjugating enzyme UFC1 (E2), and likely the UFM1-ligase E3 enzyme UFL1.

Gene References Into Functions

Crystal structure analysis of the UFM1 complex with a catalytically inactive UfSP (UFM1-specific protease) mutant reveals that, in addition to the three terminal residues of UFM1 (Pro88-Arg89-Asp90), corresponding to P6-P4 positions from the cleavage site, several conserved residues unique to UFM1 mediate extensive interactions with UfSP. PMID: 29251776
Biallelic variants in UBA5 demonstrate that disruption of the UFM1 cascade can lead to early-onset encephalopathy. PMID: 27545681

Database Links

KEGG: cel:CELE_ZK652.3

STRING: 6239.ZK652.3.2

UniGene: Cel.19419

Protein Families

UFM1 family

Tissue Specificity

Expressed in the intestine and head neurons.

Q&A

The ubiquitin-fold modifier 1 (UFM1) antibody is critical for studying ufmylation's role in cellular processes. Below are structured FAQs addressing key technical and methodological considerations for researchers, derived from peer-reviewed studies and antibody validation data.

Advanced Research Questions

How to resolve contradictions in UFM1 interaction data during co-immunoprecipitation (co-IP)?

Problem: Inconsistent detection of UFL1-UFC1-CDK5RAP3 complexes.
Solution:
- Use UFBP1 KO cells to test dependency on UFBP1 scaffolding .
- Verify endogenous protein levels via immunoblotting before IP .
- For weak interactions, coexpress UFBP1 to stabilize the E3 ligase complex .

How to design experiments investigating UFM1’s role in antiviral responses?

Model systems: Use Sendai virus (SenV)-infected HEK293T cells to monitor IFN-β/λ1 downregulation in UFM1 KO models .
Key readouts:
- RNA-seq analysis of ISGs (e.g., IFNB1, IFNL1) .
- Co-IP assays to track UFL1-14-3-3ε-RIG-I complex formation during infection .

Pathway Component	Methodological Consideration
RIG-I activation	Confirm ufmylation via UFM1G82A mutant trapping
14-3-3ε interaction	Deplete 14-3-3ε to test UFL1-RIG-I binding dependency

Methodological Best Practices

How to troubleshoot low signal in UFM1 western blots?

Cause: Transient ufmylation states or protease degradation.
Fix:
- Treat cells with proteasome inhibitors (e.g., MG132) pre-lysis .
- Use UFSP2 KO cells to accumulate di-ufmylated substrates (e.g., RPL26) .

What controls are essential for UFM1 functional studies?

Positive: Anisomycin-treated cells to induce ribosomal stress and ufmylation .
Negative: UFM1/UFL1 KO cells + rescue with catalytically dead mutants (e.g., UFL1 D129A) .

Data Interpretation Guidelines

How to distinguish artifact bands from true UFM1 conjugates?

Compare migration patterns across cell types (e.g., L02 vs. U-251 cells) .
Perform λ-phosphatase treatment to rule out phosphorylation-induced mobility shifts .

Why do UFM1 levels vary across cell lines?

Key factors:
- Basal ER stress levels (higher in HT-29 and HepG2) .
- Ribosome biogenesis activity (e.g., elevated in HeLa) .

Cell Line	UFM1 Expression Profile
HEK293T	Moderate, inducible by ER stress
Mouse liver	High constitutive levels

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ufm-1 Antibody