PPARG Antibody
Description
Definition and Biological Role of PPARG Antibody
PPARG antibodies are immunological tools designed to detect and study the peroxisome proliferator-activated receptor gamma (PPARG), a nuclear hormone receptor critical for regulating adipogenesis, lipid metabolism, and glucose homeostasis . These antibodies enable researchers to investigate PPARG's role in diseases such as obesity, diabetes, atherosclerosis, and cancer .
Types and Characteristics of PPARG Antibodies
PPARG antibodies vary in clonality, host species, and applications. Key examples include:
Key Features:
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Molecular Weight Recognition: Detects PPARG isoforms (e.g., 57 kDa PPARG1 and 54 kDa PPARG2) .
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Species Reactivity: Broad reactivity across human, mouse, rat, and other species .
3.1. Mechanistic Studies
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Adipocyte Differentiation: PPARG antibodies validate its role in lipid storage and insulin sensitivity via Western blot (WB) in 3T3-L1 cell models .
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Inflammation Modulation: PPARG transrepresses NF-κB, reducing pro-inflammatory cytokines in lung injury models .
3.2. Disease Associations
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Cancer: PPARG expression correlates with tumor prognosis; high levels in gliomas and colon cancer suggest therapeutic targeting potential .
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Autoimmunity: Pparg haploinsufficiency in mice increases autoantibody production (e.g., anti-dsDNA), implicating PPARG in lupus-like pathologies .
Key Research Findings Using PPARG Antibodies
Molecular and Functional Insights
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Domain Specificity: Antibodies targeting the N-terminal (e.g., ab45036) or ligand-binding domain (e.g., PA3-821A) reveal PPARG’s structural roles in DNA binding and coactivator recruitment .
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Post-Translational Modifications: Phosphorylation at Ser84 (detected via IP) inhibits PPARG activity, impacting insulin resistance .
Clinical and Therapeutic Implications
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Metabolic Disorders: PPARG agonists (e.g., thiazolidinediones) improve insulin sensitivity, validated using PPARG antibodies in preclinical models .
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Cancer Therapy: PPARG knockdown reduces tumor growth in vitro, highlighting its role as a therapeutic target .
Challenges and Considerations
Product Specs
Peroxisome proliferators, recognized as non-genotoxic carcinogens, are believed to influence cellular processes through their interactions with specific nuclear hormone receptors known as peroxisome proliferator-activated receptors (PPARs). These receptors belong to a family of ligand-dependent intracellular proteins that regulate gene expression. Upon activation by specific ligands, PPARs bind to particular DNA sequences, thereby stimulating the transcription of target genes. Research suggests that PPARs can be activated by various peroxisome proliferators, including clofibric acid, nafenopin, and WY-14,643, as well as certain fatty acids.
The antibody solution has a concentration of 1mg/ml and is buffered in a solution of phosphate-buffered saline (PBS) at a pH of 7.4. It also contains 10% glycerol and 0.02% sodium azide as preservatives.
This antibody has undergone rigorous testing in various applications, including ELISA, Western blot analysis, Flow cytometry, and ICC/IF, to ensure its specificity and reactivity. However, it's important to note that optimal antibody dilutions may vary depending on the specific application, and titration experiments are recommended for each investigation to determine the most effective working concentration.
Peroxisome proliferator-activated receptor gamma, PPAR-gamma, PPARG, NR1C3, PPARG1, PPARG2.
PPARG antibody was purified from mouse ascitic fluids by protein-A affinity chromatography.
PAT4F5AT
Anti-human PPARG mAb, is derived from hybridization of mouse F0 myeloma cells with spleen cells from BALB/c mice immunized with a recombinant human PPARG protein 209-477 amino acids purified from E. coli.
Mouse IgG1 heavy chain and k light chain.
Q&A
How do researchers validate PPARG antibody specificity for immunohistochemistry (IHC)?
Validation requires a multi-step approach:
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Knockout/Knockdown controls: Compare staining in PPARG-knockout models (e.g., CRISPR-modified 3T3-L1 cells) with wild-type samples .
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Western blot correlation: Confirm antibody detects a single band at 50–60 kDa in lysates from PPARG-expressing tissues (e.g., adipose tissue) .
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Blocking peptide assays: Pre-incubate antibody with immunogen peptide (e.g., Human PPARG aa 1–150) to confirm signal loss.
Example validation data from recent studies:
What are the optimal experimental conditions for PPARG western blotting?
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Lysis buffer: Use RIPA buffer with 1% SDS to solubilize nuclear fractions .
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Gel percentage: 10% SDS-PAGE resolves the 50–60 kDa band effectively .
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Blocking: 5% BSA in TBST reduces non-specific binding compared to skim milk .
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Antibody dilution: Start with 1:1,000 for monoclonal (e.g., ab41928) or 1:500 for polyclonal antibodies .
Critical note: PPARG undergoes post-translational modifications (e.g., phosphorylation at Ser84) that may cause mobility shifts. Always include a positive control like differentiated 3T3-L1 adipocytes .
How to resolve contradictory PPARG localization data between nuclear and cytoplasmic staining?
Contradictions often arise from:
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Isoform differences: PPARG1 (ubiquitous) vs. PPARG2/3 (adipose-specific) . Use isoform-specific antibodies (e.g., ab41928 for γ2) .
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Pathological contexts: Cytoplasmic PPARG accumulation occurs in 18% of papillary thyroid carcinomas . Validate with subcellular fractionation followed by Western blot.
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Antibody cross-reactivity: Polyclonal antibodies (e.g., TA326810) may detect mitochondrial proteins in metabolically active tissues. Confirm with mass spectrometry .
Case study: In thyroid cancer research, cytoplasmic PPARG staining was initially dismissed as artifactual but later linked to truncated isoforms through RNA sequencing correlation .
What methodologies optimize PPARG detection in co-immunoprecipitation (Co-IP) assays?
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Crosslinker choice: Use DSP (dithiobis[succinimidyl propionate]) for transient PPARG-RXRα interactions .
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Lysis conditions: 1% digitonin preserves PPARG-RXRα complexes better than Triton X-100 .
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Pre-clearing: Incubate lysates with Protein A/G beads for 1 hr before adding primary antibody .
Key finding from B-cell studies: PPARG forms ligand-dependent complexes with RXRα during antibody production. Co-IP with 15d-PGJ2 (1 μM) increases complex stability by 2.3-fold .
What controls are essential when comparing PPARG expression across species?
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Ortholog validation:
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Negative controls:
Table: Species cross-reactivity of common antibodies
| Antibody ID | Host | Reactivity | Key Application |
|---|---|---|---|
| ab41928 | Mouse | Human, Mouse, Rat (WB/IHC) | Nuclear staining |
| 16643-1-AP | Rabbit | Human, Mouse, Rat, Zebrafish | ChIP-seq |
| TA326810 | Rabbit | Human, Mouse (ICC only) | Co-IP |
How to address non-specific bands in PPARG Western blots?
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Band pattern analysis:
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Alternative approach: Combine IP with WB using monoclonal antibodies (e.g., A3409A) to increase specificity.
Data from proteomics studies:
Non-specific bands at 45 kDa in adipocyte lysates were identified as PPARG2 isoforms through peptide mass fingerprinting .
What novel methods are advancing PPARG research in single-cell analysis?
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CITE-seq: Pairing oligonucleotide-conjugated PPARG antibodies (e.g., TotalSeq™-B) with transcriptomics in adipose stromal cells.
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Spatial proteomics: Combine IHC-validated antibodies (TA326810) with Visium Spatial Gene Expression to map PPARG in tumor microenvironments .
Recent innovation: A 2024 study used PPARG antibody barcoding to track adipocyte differentiation at single-cell resolution, revealing three distinct PPARG expression trajectories during lipogenesis .
Product Science Overview
PPARγ exists in two main isoforms: PPARγ1 and PPARγ2. PPARγ1 is expressed in various tissues, including adipose tissue, colon, and macrophages, while PPARγ2 is predominantly found in adipose tissue and the intestine . These isoforms arise from alternative splicing of the PPARG gene, which is located on chromosome 3 in humans .
PPARγ is a key regulator of adipocyte differentiation and glucose homeostasis. It controls the peroxisomal beta-oxidation pathway of fatty acids by modulating the transcription of target genes such as acyl-CoA oxidase . PPARγ activation enhances lipid uptake and adipogenesis in fat cells, thereby increasing insulin sensitivity and reducing lipotoxicity . Additionally, PPARγ plays a role in inflammatory processes and has been implicated in the pathology of diseases such as obesity, diabetes, atherosclerosis, and cancer .
Mouse anti-human PPARγ antibodies are monoclonal antibodies produced by immunizing mice with human PPARγ protein. These antibodies are used in various research applications, including Western blotting, immunohistochemistry, and flow cytometry, to detect and quantify PPARγ expression in human tissues and cells. They are valuable tools for studying the role of PPARγ in different biological processes and disease states.
PPARγ agonists, such as thiazolidinediones, are used clinically to improve insulin sensitivity in patients with type 2 diabetes. These drugs activate PPARγ, leading to enhanced glucose uptake and reduced blood glucose levels . However, PPARγ activation can also have side effects, including weight gain and fluid retention . Research is ongoing to develop more selective PPARγ modulators with fewer adverse effects.
