PKL Antibody
Description
Biochemical Characteristics and Target Specificity
PKL antibodies are polyclonal reagents produced in rabbits, targeting the human PKLR protein (UniProt ID: P30613). Key properties include:
These antibodies exhibit minimal cross-reactivity with other pyruvate kinase isoforms (e.g., PKM2) , ensuring specificity for PKLR in assays.
Applications in Research and Diagnostics
PKL antibodies are widely used in multiple experimental workflows:
Key Applications
These reagents have been employed to study PKLR’s role in metabolic diseases and cancer, with protocols available for tissue-specific antigen retrieval (e.g., TE buffer pH 9.0 for IHC) .
Validation and Quality Control
Antibody validation is critical to ensure specificity:
-
Genetic Controls: Knockout (KO) cell lines confirm absence of off-target binding .
-
Orthogonal Methods: Correlation with mass spectrometry or RNA-seq data enhances reliability .
-
Multi-Antibody Validation: Concordance across independent PKLR antibodies (e.g., Proteintech 22456-1-AP vs. R&D Systems AF8519) supports specificity .
The YCharOS study highlighted that 50–75% of commercial antibodies pass rigorous validation, with recombinant antibodies outperforming polyclonals in reproducibility . Vendors like Proteintech and Sigma-Aldrich provide application-specific validation data, including KO validation in hepatic models .
Research Implications
PKL antibodies have advanced studies on:
-
Metabolic Disorders: PKLR mutations cause pyruvate kinase deficiency, a hereditary hemolytic anemia .
-
Cancer Metabolism: Overexpression in hepatic and breast cancers correlates with glycolytic flux .
-
Chromatin Remodeling: In plants, PKL (PICKLE) antibodies elucidate roles in histone modification and gene silencing .
Future directions include leveraging recombinant antibodies for enhanced reproducibility and developing PKLR-targeted therapies for metabolic diseases .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- The antagonistic functions of FIS1 and PKR2 in modulating endosperm development mirror those of PICKLE (PKL) and CURLY LEAF (CLF), which antagonistically regulate root meristem activity. PMID: 28155248
- Pickle plays a significant role in vegetative growth and gibberellin signaling. PMID: 28057895
- Plants lacking the CHD3 remodeler, PICKLE, exhibit various reproductive defects. These phenotypes stem from the loss of PICKLE in the maternal sporophyte. PMID: 27075727
- Research suggests that transcriptional regulation of LFY at the chromatin level by PKL may partially account for the late-flowering phenotype observed in pkl mutants. PMID: 27056257
- EPP1 works in conjunction with SPA1 to repress seedling de-etiolation. PMID: 23733056
- PKL interacts physically with HY5 to directly regulate hypocotyl cell elongation by repressing trimethylation of histone H3 Lys 27 (H3K27me3) on the regulatory regions of several cell elongation-related genes in response to changing light conditions. PMID: 23314848
- These studies demonstrate that subfamily II CHD proteins in plants, such as PICKLE, retain ATP-dependent chromatin remodeling activity. PMID: 23128324
- CKH1/EER4/AtTAF12b and CKH2/PKL may collaborate on cytokinin-regulated genes. PMID: 21357580
- PKL plays a role in maintaining chromatin homeostasis. PMID: 22452853
- Loss of PKL function hinders root growth in Arabidopsis. PMID: 21441433
- PKL is continuously required after germination to repress the expression of PHERES1, a type I MADS box gene normally expressed during early embryogenesis in wild-type plants. PMID: 16359393
- PKL-dependent repression of embryonic gene expression extends to late-embryogenesis genes and is associated with changes in chromatin. PMID: 17892443
- PKL promotes histone H3 methylation in both germinating seedlings and adult plants, but a connection between PKL-dependent expression and acetylation levels has not been identified. PMID: 18539592
- PKL and PKR2 are essential for the expression of many genes that are repressed by PcG proteins. PMID: 19680533
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with antibody-based methodologies, synthesized from interdisciplinary insights across protein detection, antibody engineering, and mechanistic studies:
Advanced Research Questions
How do I resolve discrepancies between in vitro and in vivo antibody efficacy?
-
Analytical framework:
| Format | Expression Yield (mg/L) | Monomer Content (%) | Clinical Relevance |
|---|---|---|---|
| Chimeric | 2.5 | 92 | Early-stage diagnostics |
| Humanized (VH+) | 75 | >99.5 | Therapeutic candidates |
-
Investigate tissue-specific post-translational modifications (e.g., glycosylation) using mass spectrometry .
What computational tools can predict antibody-antigen interfaces to guide rational engineering?
-
Pipeline:
How can antibody specificity be leveraged to dissect metabolic pathway crosstalk in disease models?
-
Case study: BRCA1 overexpression downregulates PKM2 via PI3K/AKT, attenuating glycolysis in breast cancer :
-
Apply phospho-specific antibodies to map kinase activation cascades.
-
Pair with siRNA knockdowns to isolate antibody-dependent metabolic effects.
-
Methodological Challenges
What strategies improve antibody stability in long-term biosensor applications?
-
Engineering solutions:
How do I validate antibody performance in multiplexed proteomic assays?
-
Validation matrix:
Data Interpretation
How should I contextualize conflicting reports of antibody therapeutic efficacy?
-
Critical factors:
