Customize tra-1 Antibody

Description

Research Applications

Tra-1 antibodies are critical tools in stem cell biology and cancer research.

Application	Method	Use Case
Flow Cytometry	Cell surface staining	Identifying undifferentiated hESCs/iPSCs
Immunohistochemistry	Tissue section staining	Detecting TRA-1+ cells in gastric cancer
Immunoprecipitation	Podocalyxin isolation	Characterizing glycan-protein interactions
Therapeutic Targeting	Antibody-drug conjugates (ADCs)	Eliminating chemoresistant cancer cells

Notable Studies:

Stem Cell Validation: BG01V hESCs stained with Tra-1-81 show membrane-specific binding .
Cancer Targeting: TRA-1-60-targeted ADCs (e.g., Bstrongomab) reduce tumor ascites in gastric cancer models .

Clinical Relevance in Oncology

TRA-1+ cells are enriched in chemoresistant tumors and correlate with poor prognosis.

Cancer Type	TRA-1+ Cell Role	Therapeutic Implication
Gastric Cancer	Invasive front cells with stress resistance	Targeting TRA-1+ cells reduces ascites
Pancreatic Cancer	Chemoresistant subpopulation	UGT1A10 inhibition via Cymarin enhances chemosensitivity
Follicular Lymphoma	Rituximab-resistant cells	TRA-1+ cells exhibit tumor-initiating capacity
Prostate Cancer	Metastatic tumor-initiating cells	TRA-1+ cells correlate with progression

Mechanistic Insights:

Chemoresistance: TRA-1+ cells upregulate drug transporters (e.g., ABCB1) and exhibit enhanced survival pathways .
Biomarker Potential: TRA-1 expression predicts chemoresistance and short survival in pancreatic cancer .

Molecular Pathways Regulating TRA-1 Expression

The UGT1A10 enzyme is critical for TRA-1 glycan synthesis and chemoresistance:

Molecule	Function	Targeted Therapy	Source
UGT1A10	Glycosylates podocalyxin	Cymarin (inhibitor) eliminates TRA-1+ cells
Podocalyxin	Carries TRA-1 epitopes	ADCs disrupt tumor-stromal interactions

Product Specs

Buffer

**Preservative:** 0.03% Proclin 300
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

tra-1 antibody; her-2 antibody; Y47D3A.6 antibody; Sex-determining transformer protein 1 antibody; Hermaphrodization of XO animals protein 2 antibody

Target Names

tra-1

Uniprot No.

P34708

Target Background

Function

TRA-1 antibody plays a critical role in regulating sexual phenotype in organisms. It serves as a terminal global regulator within a well-defined cascade of sex-determining genes, promoting female development. TRA-1 interacts with TRA-2 to facilitate spermatogenesis. It further promotes spermatogenesis through the Tip60 HAT complex and by controlling the expression of genes essential for male development, such as *fog-3*. Its association with chromatin and the *fog-3* promoter requires WDR-5.1 and potentially WDR-5.2. In collaboration with TRR-1, TRA-1 activates the *fog-3* gene, ultimately determining sperm/oocyte cell fate. In hermaphrodites, TRA-1 binds to an intronic regulatory site within the *ceh-30* gene, inhibiting its transcription and preventing the survival of CEM (cephalic male) sensory neurons. It represses the expression of the transcription factor DMD-3 in hermaphrodites, governing the timing and extent of male tail tip morphogenesis. TRA-1 also participates in controlling the sex-specific differentiation of PHC sensory neurons and suppresses the development of male-specific morphological features.

Database Links

KEGG: cel:CELE_Y47D3A.6

STRING: 6239.Y47D3A.6a

UniGene: Cel.9756

Protein Families

GLI C2H2-type zinc-finger protein family

Subcellular Location

Cytoplasm. Nucleus.

Tissue Specificity

Expressed in intestine and gonads (at protein level).

Q&A

Here’s a structured collection of advanced FAQs for researchers working with TRA-1 antibodies, synthesized from peer-reviewed studies and technical protocols:

How do TRA-1-60 and TRA-1-81 antibodies validate pluripotency in human stem cells?

Methodology:
- Use flow cytometry (10 µg/mL antibody concentration, IgM isotype controls) to detect surface antigens on undifferentiated embryonic stem cells (e.g., BG01V line) .
- Confirm localization via immunofluorescence (3-hour room-temperature incubation with NorthernLights™ 557 secondary antibodies) .
- Cross-validate with SSEA-3/4 positivity and SSEA-1 negativity .
Key Data:

TRA-1-81 binds a keratan sulfate-modified podocalyxin epitope distinct from TRA-1-60, requiring β1,3-linked type 1 lactosamine extensions .

What controls are critical for TRA-1 antibody specificity in Western blotting?

Experimental Design:
- Include lysates from NTERA-2 embryonal carcinoma cells (positive control) and differentiated somatic cells (negative control) .
- Use enzymatic digestion (neuraminidase) to confirm carbohydrate-dependent epitopes .
Common Pitfalls:
Non-specific bands at 200–400 kDa may appear due to glycosylation variability; validate with siRNA knockdown of podocalyxin .

How do TRA-1-60/81+ cells drive chemoresistance in pancreatic cancer?

Mechanistic Insights:
- TRA-1-60/81+ cells exhibit upregulated UGT1A10, which conjugates glucuronic acid to chemotherapeutics (e.g., gemcitabine), enhancing drug efflux .
- In vivo validation: Xenograft models show 3.2-fold enrichment of TRA-1-60+ cells post-chemotherapy (p < 0.001) .
Therapeutic Strategy:

Target Intervention Outcome
UGT1A10 Cymarin (IC₅₀: 0.8 µM) Reduces TRA-1-60/81+ cells by 74% in PDAC models

Target	Intervention	Outcome
UGT1A10	Cymarin (IC₅₀: 0.8 µM)	Reduces TRA-1-60/81+ cells by 74% in PDAC models

How to resolve contradictory TRA-1 epitope data across cancer types?

Analytical Framework:
- Tissue-specific glycosylation: TRA-1-81 recognizes sialofucosylated podocalyxin in metastases but not primary tumors .
- Antibody cross-reactivity: Perform glycan array screening (500+ structures) to confirm binding to Galβ1-3GlcNAcβ1-3Galβ1-4GlcNAc .
- Context-dependent expression:
  - 12% of pancreatic tumors vs. <5% of inflamed tissues express TRA-1-60 .
  - No detection in normal tissues .

What experimental models best capture TRA-1 dynamics during differentiation?

Optimized Workflow:
- 2D vs. 3D cultures: Use Matrigel-embedded organoids to mimic in vivo antigen retention (12-day differentiation assay) .
- Retinoic acid titration: 1 µM RA reduces TRA-1-81 expression by 83% within 72 hours (qPCR validation) .
- Single-cell RNA-seq: Cluster cells by PODXL expression to correlate with TRA-1 antibody staining .

Technical Comparison Table

Parameter	TRA-1-60	TRA-1-81
Epitope	Sialylated keratan sulfate	Non-sialylated β1,3-galactosylated KS
Neuraminidase sensitivity	Enhanced binding	No effect
Cancer relevance	32.7% vs. 49.9% positivity in long vs. short PDAC survival	Aggressive breast/prostate cancer

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tra-1 Antibody