Customize MT1A Antibody

Description

Overview of MT1A Antibodies

MT1A antibodies are primarily polyclonal or monoclonal reagents designed to detect Metallothionein 1A protein in various experimental systems. They are validated for techniques such as immunohistochemistry (IHC), immunofluorescence (IF), Western blot (WB), and enzyme-linked immunosorbent assay (ELISA).

Key Features	Details
Host Species	Rabbit (polyclonal), Mouse (monoclonal)
Reactivity	Human, Mouse, Rat, Rabbit, Bovine
Applications	IHC, IF, WB, ELISA, Flow Cytometry
Conjugates	Unconjugated, HRP, FITC, Biotin
Molecular Weight Target	6 kDa (observed and calculated)

Applications of MT1A Antibodies

MT1A antibodies are employed in diverse experimental contexts, with specific dilutions and protocols optimized for each technique.

Immunohistochemistry (IHC)

Human Tissue Detection: Positive staining in kidney and liver tissues using TE buffer (pH 9.0) or citrate buffer (pH 6.0) for antigen retrieval .
Dilution: 1:50–1:500 .

Immunofluorescence (IF)

Cell Line Detection: HeLa cells show cytoplasmic/nuclear localization using Alexa Fluor® 488/594 conjugates .
Dilution: 1:200–1:800 .

Western Blot (WB)

Predicted Band: 6 kDa .
Sample Types: HeLa lysates, rabbit liver lysates .
Dilution: 1/1000–1/5000 .

ELISA

Quantitative Measurement: Bovine MT1A antibody pair sets use sandwich ELISA for serum, plasma, or tissue lysates .
Sensitivity: Optimized for high-throughput assays .

Comparative Table of MT1A Antibodies

Product	Host	Reactivity	Applications	Dilution
27115-1-AP (Proteintech)	Rabbit	Human	IHC, IF, ELISA	IHC: 1:50–1:500; IF: 1:200–1:800
16713-1-AP (Proteintech)	Rabbit	Human, Mouse, Rat	ELISA	N/A
UC1MT (Abcam)	Mouse	Human, Rabbit	WB, IF, Flow Cytometry	WB: 1/1000; IF: 1–10 μg/ml
Bovine Pair Set (Creative Biolabs)	Goat/Rabbit	Bovine	ELISA	Capture: 1:100–1:500; Detection: 1:500–1:2000

Role in Cancer and Oxidative Stress

Oncogenic Potential: MT1A overexpression is linked to tumor progression, angiogenesis, and chemotherapy resistance in cancers (e.g., lung, prostate, cervical) .
Oxidative Stress Response: MT1A upregulation protects cells against reactive oxygen species and heavy metal toxicity .

Diagnostic and Therapeutic Relevance

Biomarker Potential: Serum MT1A levels correlate with tumor staging and prognosis, though species-specific antibodies are required for accurate measurement .
Immunotherapy: MT1A antibodies aid in studying PD-1/PD-L1 interactions and T-cell responses in cancer immunotherapy .

Antigen Retrieval for IHC

Buffer Options: TE (pH 9.0) or citrate (pH 6.0) for human tissues .
Fixation: 4% PFA for IF/ICC .

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

MT1A antibody; LSR4 antibody; At1g07600 antibody; F22G5_33 antibody; F24B9.34 antibody; Metallothionein-like protein 1A antibody; MT-1A antibody; MT-2 antibody; MT-Q antibody; Protein LIGHT STRESS-REGULATED 4 antibody

Target Names

MT1A

Uniprot No.

P43392

Target Background

Function

Metallothioneins are proteins rich in cysteine residues, which bind to various heavy metals. They are believed to function as metal chelators, particularly for copper (Cu) and zinc (Zn). MT1A plays a significant role in copper homeostasis within plant roots when exposed to elevated copper concentrations. It collaborates with the phytochelatin synthase PCS1 to protect plants from the toxic effects of copper and cadmium (Cd). MT1A also contributes to copper homeostasis by facilitating the remobilization of copper from senescing leaves. This copper mobilization from internal sources is essential for seed development. Furthermore, MT1A confers tolerance to cadmium and participates in cadmium and zinc homeostasis.

Database Links

KEGG: ath:AT1G07600

STRING: 3702.AT1G07600.1

UniGene: At.23699

Protein Families

Metallothionein superfamily, Type 15 family

Tissue Specificity

Expressed in phloem and mesophyll cells of leaves, vascular tissues of cotyledons, sepals and petals. Expressed in anthers. Expressed in root endodermis and at lower levels in cortex of mature region of roots.

Q&A

Basic Research Questions

How to validate MT1A antibody specificity in cross-reactive experimental models?

Validation requires a multi-step approach:

Knockout Controls: Use MT1A-deficient cell lines or tissues (e.g., CRISPR-edited models) to confirm absence of signal .
Western Blotting: Verify antibody binds to the expected ~6.1 kDa band under non-reducing conditions .
Immunohistochemistry (IHC): Compare staining patterns with known MT1A-positive/negative tissues (e.g., liver vs. brain) .

What factors influence MT1A antibody performance in different applications?

Application	Key Considerations
Western Blot	Use Tris-Tricine gels for optimal separation of low-molecular-weight proteins .
IHC	Antigen retrieval with citrate buffer (pH 6.0) enhances epitope accessibility .
ELISA	Pre-adsorb recombinant MT1A to confirm linearity in standard curves .

How to resolve discrepancies in MT1A detection across studies?

Sample Preparation: Metallothioneins are cysteine-rich; avoid β-mercaptoethanol to prevent protein aggregation .
Antibody Clonality: Polyclonal antibodies may detect isoforms (e.g., MT1 vs. MT2), while monoclonals require rigorous isoform validation .

Advanced Research Questions

How do MT1A SNPs (e.g., rs11076161) impact antibody-based quantification?

The rs11076161 (A > G) polymorphism alters MT1A expression levels, requiring genotype stratification:

Genotype	Median MT2 Level (μg/g)	p-Value
GG (n=37)	0.194 vs. 0.227	0.941
AG (n=31)	0.237 vs. 0.239	0.913
AA (n=4)	0.174 vs. 0.197	-
Data adapted from endometrial tissue study

Methodological Recommendation:

Genotype samples using TaqMan assays (e.g., Thermo Fisher C_1402094_10) before quantitative analysis.

What strategies address MT1A’s role in immune regulation during functional studies?

Co-culture Systems: Use MT1A-overexpressing macrophages + T cells to assess cytokine modulation (e.g., IL-10/IL-12 balance) .
Metal Chelation: Deplete zinc with TPEN to isolate MT1A’s metal-independent immunomodulatory effects .

How to reconcile conflicting data on MT1A’s pro- vs. anti-inflammatory roles?

Context-Dependent Analysis:
- Pro-inflammatory: Measure MT1A-NFκB interaction in LPS-treated monocytes .
- Anti-inflammatory: Quantify MT1A-mediated ROS scavenging in colitis models .
Single-Cell RNA Sequencing: Resolve cell-type-specific MT1A expression in heterogeneous tissues .

Data Contradiction Analysis

How to interpret variability in MT1A expression across cancer studies?

Factor	Impact	Resolution Strategy
Tumor Stage	Early vs. late-stage tumors show inverse MT1A trends	Stratify cohorts by TNM classification .
Metal Exposure	Cadmium upregulates MT1A, confounding disease associations	Control for environmental metal levels via ICP-MS .

Why do some MT1A antibodies fail in primate models despite human reactivity?

Epitope Divergence: Non-conserved residues in primate MT1A (e.g., position 12: human Lys → primate Glu) .
Solution: Use antibodies raised against primate-specific MT1A peptides (e.g., residues 30-45) .

Technical Troubleshooting

How to optimize MT1A detection in multiplex assays?

Cross-Reactivity Check: Pre-incubate antibodies with MT2A to block shared epitopes .
Signal Amplification: Employ tyramide-based systems for low-abundance MT1A in FFPE tissues .

What controls are essential for MT1A knockout validation?

Control Type	Purpose
Isoform-Specific KO	Exclude MT2A/MT3 compensatory effects
Metal Challenge	Verify loss of zinc-induced MT1A response
Baseline ROS	Confirm altered oxidative stress in KO models

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MT1A Antibody