ZNF230 Antibody
Description
Structure and Function of ZNF230 Antibody
ZNF230 Antibody is designed to detect the ZNF230 protein, which belongs to the zinc finger protein (ZNF) family. These proteins are characterized by zinc-coordinating motifs that stabilize their structure, enabling interactions with DNA, RNA, or other proteins .
Applications in Research
ZNF230 Antibody is validated for immunohistochemistry (IHC), Western blot (WB), ELISA, and immunoprecipitation (IP). Key applications include:
-
Transcriptional Regulation Studies: ZNF230’s role in RNA polymerase II-mediated transcription and gene expression control .
-
Genome Stability Research: Investigating ZNF230’s potential involvement in DNA repair pathways, as zinc finger proteins are critical for genome integrity .
-
Disease Modeling: Analyzing ZNF230’s dysregulation in cancers and developmental disorders .
IHC and WB Protocols
| Application | Recommended Dilution | Sample Type | Source |
|---|---|---|---|
| IHC | 1:20–1:200 | Paraffin-embedded tissues | |
| WB | 1:500–1:2000 | Mouse liver, human testis |
Functional Insights
-
Transcriptional Regulation: ZNF230 may recruit co-repressors or activate transcription through interactions with chromatin-modifying enzymes .
-
Testicular Function: Targeted disruption of Znf230 in mice led to infertility, highlighting its critical role in spermatogenesis .
-
Genome Stability: Zinc finger proteins like ZNF230 are implicated in DNA double-strand break repair, though direct evidence for ZNF230 remains under investigation .
Tissue Expression
ZNF230 exhibits testis-enriched expression in humans, as confirmed by the Human Protein Atlas . It is also detected in other tissues, including the brain, liver, and kidney, albeit at lower levels .
Available Antibodies and Comparisons
A range of ZNF230 Antibodies are commercially available, differing in clonality, conjugation, and validated applications. Below is a comparison:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- The ZNF230 gene has been implicated in azoospermia, with the A316G mutation potentially correlated with serum FSH levels. PMID: 15952109
- Transcriptional control plays a significant role in regulating ZNF230 expression, with a common mechanism governing its basal transcription. PMID: 18584306
Q&A
How should researchers validate ZNF230 antibody specificity for Western blot or immunoprecipitation?
Validation requires a multi-step approach:
-
Knockout Controls: Use tissues or cell lines lacking ZNF230 (e.g., Znf230 KO mouse testes ) to confirm absence of signal in Western blot (WB) or immunoprecipitation (IP). For example, the 26-kDa band observed in wild-type mice disappears in KO models .
-
Immunogen Alignment: Verify that the antibody’s immunogen sequence (e.g., residues 85–249 for PCRP-ZNF230-3C11 ) overlaps with known functional domains (e.g., the RING finger motif at residues 218–230 ).
-
Cross-Reactivity Screening: Test species specificity using samples from human, mouse, and other models. PCRP-ZNF230-3C11 is confirmed for human reactivity only .
Table 1: Validation Parameters for ZNF230 Antibodies
What are the recommended applications for ZNF230 antibodies in basic research?
The PCRP-ZNF230 clones are optimized for:
-
Immunoprecipitation: Use mild lysis buffers (e.g., RIPA with protease inhibitors) to preserve protein-protein interactions. Pre-clear lysates with protein A/G beads to reduce non-specific binding .
-
Microarray Analysis: Leverage transcriptomic data from Znf230 KO models to identify downstream targets (e.g., Kctd14, reduced 10-fold in KO mice ).
-
Subcellular Localization: In wild-type mice, ZNF230 localizes to spermatogonial nuclei and sperm tails , suggesting immunofluorescence (IF) protocols should include nuclear/cytoskeletal fractionation.
How should storage conditions be optimized for ZNF230 antibodies?
-
Short-Term: Aliquot into 20 µL volumes and store at 4°C for ≤2 weeks .
-
Long-Term: Add glycerol (1:1 ratio) and freeze at -80°C. Avoid freeze-thaw cycles, which destabilize IgG2b/IgG1 conformations .
-
Activity Monitoring: Run a WB with a positive control (e.g., wild-type testis lysate) every 6 months to confirm epitope integrity .
How can epitope mapping resolve discrepancies in ZNF230 antibody performance?
The PCRP-ZNF230-1E1 antibody maps to residues 85–249 , while KO studies show that the RING domain (residues 218–230) is critical for function . If an antibody fails to detect ZNF230 in a functional assay:
-
Epitope Truncation: Express truncated ZNF230 variants (e.g., Δ85–249) to test antibody binding.
-
Competition Assays: Co-incubate antibodies with recombinant immunogen peptides (10–100 µg/mL) to block binding .
-
Structural Modeling: Use AlphaFold-predicted ZNF230 structures to assess epitope accessibility in different conformational states .
How do transcriptomic findings in Znf230 KO mice inform antibody-based mechanistic studies?
Despite normal spermatogenesis in Znf230 KO mice, microarray data revealed dysregulation of four transcripts :
-
Upregulated: 4921513I08Rik (2.1-fold).
-
Downregulated: Kctd14 (10-fold), 4930448A20Rik, 4931431B13Rik.
To link ZNF230 antibody data to these findings:
-
Co-IP/MS: Identify ZNF230-binding partners (e.g., Kctd14 regulators) using immunoprecipitation followed by mass spectrometry.
-
ChIP-seq: Use ZNF230 antibodies to map DNA-binding sites and correlate with differentially expressed genes.
What experimental strategies address contradictory phenotypic data in ZNF230 studies?
The absence of fertility defects in Znf230 KO mice contrasts with its testis-enriched expression. To reconcile this:
-
Conditional Knockout Models: Use cell-specific Cre drivers (e.g., Stra8-Cre for germ cells) to bypass compensatory mechanisms.
-
High-Throughput Phosphoproteomics: Quantify post-translational modifications in KO vs. wild-type spermatozoa.
-
Multi-Antibody Validation: Compare results from PCRP-ZNF230-3C11 (IgG2b) and PCRP-ZNF230-1E1 (IgG1) to rule out isoform-specific artifacts .
How can ZNF230 antibodies be integrated into multiplexed assays for systems biology?
-
Spatial Transcriptomics: Combine immunofluorescence (IF) with RNA-ISH to correlate ZNF230 protein levels with Kctd14 mRNA in testis sections.
-
Flow Cytometry Panels: Use ZNF230 antibodies with germ cell markers (e.g., VASA, CREM) to stratify spermatogenic stages.
-
Machine Learning: Train classifiers on antibody-derived data (e.g., IF intensity, IP abundance) to predict transcriptional outcomes .
Methodological Best Practices
-
Controls for IP/WB: Include KO tissues, isotype-matched IgG, and peptide-blocked antibody samples.
-
Quantitative Thresholds: Define positivity thresholds for IF (e.g., >2-fold signal vs. KO).
-
Data Triangulation: Cross-validate antibody findings with CRISPRi knockdown or mRNA-seq.
