Customize ubxn-4 Antibody

Description

UBXN4 Antibody Overview

The UBXN4 antibody targets the UBXN4 protein (UniProt ID: Q92575), encoded by the UBXN4 gene (NCBI Gene ID: 23190). This protein is also known as UBXD2 or UBX domain protein 4. Key features include:

Property	Details
Host Species	Rabbit (IgG)
Reactivity	Human, Mouse, Rat
Observed MW	64–68 kDa
Applications	WB, IHC, IF/ICC, IP, ELISA
Immunogen	UBXD2 fusion protein (Ag13440)
Key Domains	UBX domain, transmembrane region

Biological Role of UBXN4

UBXN4 is an integral membrane protein localized to the endoplasmic reticulum (ER). It functions as an adaptor in ERAD by recruiting VCP (valosin-containing protein) and UBQLN1 to facilitate the degradation of misfolded proteins . Key roles include:

ER-Mitochondria Contact Regulation: Modulates lipid saturation and composition at ER-mitochondria junctions .
Aggresome Formation: Assists in organizing protein aggregates for degradation via the p97-UBXN1 complex .
Nicotinic Receptor Trafficking: Binds α3/α4 nicotinic acetylcholine receptor subunits to regulate surface expression .

Antibody Validation and Applications

The UBXN4 antibody (e.g., Proteintech 21052-1-AP) has been validated across multiple platforms:

Key Validation Data

Application	Results
Western Blot (WB)	Detects endogenous UBXN4 in HEK-293 cells, mouse/rat brain tissue .
Immunoprecipitation	Confirmed interaction with VCP and UBQLN1 in mouse brain lysates .
Immunofluorescence	Localizes UBXN4 to ER and cis-Golgi compartments in HEK-293 cells .
Immunohistochemistry	Staining observed in human colon cancer tissues .

Table 1: Key Studies on UBXN4

Study	Findings	Citation
ERAD Mechanism	UBXN4 stabilizes interactions between VCP and UBQLN1 to enhance ERAD efficiency .
Mitochondrial Regulation	UBXN4-containing complexes alter lipid composition at ER-mitochondria contact sites .
Nicotinic Receptor Trafficking	UBXN4 knockdown reduces α3β2 nicotinic receptor surface expression in neurons .
Proteasome Coupling	UBXN4 deficiency correlates with impaired ubiquitination-proteasome coupling in cardiac cells .

Recommended Dilutions

Application	Dilution Range
WB	1:2,000–1:10,000
IHC	1:50–1:500
IF/ICC	1:50–1:500

Functional Insights

Ubiquitination Control: UBXN4 reduces ubiquitination of α3 nAChR subunits, preventing proteasomal degradation .
Pathological Relevance: Dysregulation of UBXN4 is implicated in inclusion body myopathy and neurodegenerative disorders .

Protocols and Troubleshooting

Antigen Retrieval: For IHC, use TE buffer (pH 9.0) or citrate buffer (pH 6.0) .
Storage: Stable at -20°C in PBS with 0.02% sodium azide and 50% glycerol .

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

ubxn-4 antibody; ZK353.8 antibody; UBX domain-containing protein 4 antibody; Erasin antibody

Target Names

ubxn-4

Uniprot No.

P34631

Target Background

Function

Likely functions as an adapter protein for the ATPases CDC-48.1 and/or CDC-48.2, thereby conferring substrate specificity. It may participate in the endoplasmic reticulum-associated protein degradation (ERAD) pathway, potentially serving as a scaffold to recruit both UBL-1 and CDC-48.1 and/or CDC-48.2 to the ER during ERAD.

Database Links

KEGG: cel:CELE_ZK353.8

STRING: 6239.ZK353.8.3

UniGene: Cel.9571

Subcellular Location

Endoplasmic reticulum membrane; Peripheral membrane protein.

Tissue Specificity

Expressed in the germline.

Q&A

Here’s a structured FAQ for researchers studying UBXN4 antibody applications in academic contexts, synthesized from peer-reviewed studies and technical methodologies:

Advanced Experimental Design

How to resolve contradictory data on UBXN4’s role in mitophagy versus aggresome formation?

Contextual Factors:

Condition UBXN4 Function Key Interactors Reference
Proteasome Inhibition Aggresome disassembly ATAD1, PLAA
Hypoxia Mitophagy regulation (putative) UBR4, TRIM25
Method: Use tissue-specific knockout models (e.g., muscle vs. liver) and condition-dependent ubiquitination assays (K48/K63 linkage analysis) .

Condition	UBXN4 Function	Key Interactors	Reference
Proteasome Inhibition	Aggresome disassembly	ATAD1, PLAA
Hypoxia	Mitophagy regulation (putative)	UBR4, TRIM25

What functional assays confirm UBXN4’s role in protein quality control?

Stepwise Protocol:
- Induce protein aggregation (e.g., 24h treatment with 10µM MG132) .
- Quantify insoluble ubiquitinated proteins via sequential detergent extraction (RIPA buffer → urea/SDS) .
- Compare UBXN4-KO vs. wild-type cells using Proteostat staining .

How to optimize UBXN4 antibody for proximity ligation assays (PLA) in fixed tissues?

Fixation: 4% PFA (avoid methanol fixation to preserve epitopes).
Validation: Pair with validated primary antibodies against binding partners (e.g., PLAA, ATAD1) .
Troubleshooting: High background? Pre-clear tissues with Fab fragments against IgG Fc regions .

Data Interpretation & Technical Challenges

Why does UBXN4 show variable migration on SDS-PAGE across studies?

Key Variables:
- Post-translational modifications (e.g., phosphorylation at Ser65 detected by Phos-tag gels) .
- Tissue-specific isoform expression (UniProt: Q9BZD5 has 3 splice variants).
Solution: Treat lysates with λ-phosphatase and repeat immunoblot .

How to address cross-reactivity with UBXD family members in IP-MS?

Strategy:
- Perform competitive elution with UBX domain peptides .
- Use CRISPR-Cas9 engineered UBXN4-KO cells as negative controls .

Critical Methodological Notes

Ubiquitination Assays: For studying UBXN4’s role in K48/K63-linked chains, use linkage-specific antibodies (e.g., Millipore 05-1307/1308) .
Aggresome Studies: Combine proteasome inhibition (e.g., 1µM Bortezomib) with UBXN4 siRNA to distinguish its role from UBXN1 .

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ubxn-4 Antibody