The POK7 antibody recognizes ZBTB7B, also known as ThPOK (T-helper-inducing POZ/Kruppel-like factor), a transcriptional regulator essential for CD4+ T cell fate. This antibody has become a core tool for immunologists, particularly in studies involving thymic development, peripheral T cell differentiation, and autoimmunity models.

What Makes ZBTB7B / POK7 Important?

ZBTB7B acts as a lineage commitment switch. It inhibits CD8 cytotoxic program genes in developing thymocytes and upregulates helper lineage identity genes, driving MHC class II-restricted cells toward the CD4+ fate.

Key functional studies:

• NIH – T Cell Lineage Fate Mapping

• PubMed – ThPOK and T Cell Differentiation

• Stanford – Genetic Control of Thymocyte Maturation

Recommended Experimental Protocols Using POK7 Antibody

1. Intracellular Flow Cytometry for ZBTB7B

Since POK7 is a nuclear protein, the cell membrane must be permeabilized. Fixation with 4% paraformaldehyde followed by saponin-based buffers (e.g., BD Perm/Wash) is optimal. Detection is strongest in:

• CD4 single-positive thymocytes

• Peripheral CD4+ T cells

• Early double-positive (CD4+CD8+) subsets with intermediate expression

Protocols and examples:

• NIH – Flow Cytometry Protocols

• Yale University – Intracellular Cytokine Staining Methods

• Harvard – Flow Core Resources

2. Western Blot Detection of POK7

Prepare nuclear lysates using a buffer with protease and phosphatase inhibitors. Load 20–30 µg per well. Antibody binding usually shows a band at ~60 kDa, depending on isoform.

Refer to:

• NCBI Protocols – Transcription Factor Western Blot

• NIEHS – Protein Extraction Methods

• University of Florida – Molecular Core Labs

Bioinformatics Support for POK7 Expression

Tissue & Cell Type Distribution

• High: Thymus, spleen, lymph nodes

• Moderate: Peripheral blood mononuclear cells

• Low/Absent: Non-immune tissues

Data sources:

• Human Protein Atlas – ZBTB7B Expression

• GTEx – Gene Expression Across Tissues

• NCBI GEO – ZBTB7B Dataset Browser

Gene Regulation & Co-expression

ZBTB7B expression correlates with:

• GATA3, BCL11B, TCF7 in CD4+ cells

• RUNX3, EOMES repression in cytotoxic CD8+ lineage

Explore:

• UCSC Genome Browser – ZBTB7B

• Ensembl – ZBTB7B Gene Tree and Transcripts

• NCBI – Conserved Domains of ThPOK

Co-Staining Panels Including POK7 Antibody

When performing multi-parameter staining, include:

• Surface: CD3, CD4, CD8, CD44, CD62L

• Transcription Factors: T-bet (TBX21), GATA3, FOXP3, RUNX3

• Cytokines (in activated conditions): IL-2, IFN-γ, IL-4

Resources:

• University of Washington – Multicolor Flow Panel Design

• NIH – Multicolor Panel Optimization

• BD Biosciences – Panel Design Tools

Troubleshooting POK7 Antibody Experiments

See best practices:

• NIH – Antibody Optimization Guidelines

• CDC – Biosafety and Sample Handling

• FDA – Protein Assay Validation Standards

Reagents, Storage, and Formats

POK7 antibodies are typically available in:

• Unconjugated format for Western blot or ELISA

• Fluorophore-conjugated (e.g., Alexa Fluor 488, PE) for FACS

• HRP-labeled for chromogenic IHC

Store at –20°C for long-term or 4°C for short-term. Protect fluorophores from light.

Detailed handling tips:

• NIH ORIP – Lab Reagent Management

• CDC – Laboratory Best Practices

• University of Pennsylvania – Antibody Reagent Guide

Summary and Key Applications

The POK7 antibody is crucial for:

• T-cell lineage analysis

• Intracellular transcription factor profiling

• Autoimmune disease modeling

• Thymic education and selection studies

• Gene regulation research involving BTB-ZF proteins

It remains one of the most specific markers for CD4+ helper lineage identification and a powerful tool in immunology, hematology, and T cell biology.

For more:

• Antibody Registry – POK7/ZBTB7B Entries

• NIH RePORTER – POK7 Funding Projects

• NCBI – Latest ThPOK Research Publications