PTPRC Monoclonal Antibody,Purified

What is PTPRC and what is its significance in immunological research?

PTPRC (Protein Tyrosine Phosphatase Receptor Type C), also known as CD45, is a cell surface glycoprotein expressed on all nucleated hematopoietic cells. It functions as a critical regulator of T and B cell antigen receptor signaling. PTPRC is essential for normal immune system development and function, making it a significant target in immunological research, particularly for studies involving leukocyte identification, activation, and development. Monoclonal antibodies against PTPRC serve as valuable tools for tracking and analyzing immune cell populations and their functional states .

What are the common applications for purified PTPRC monoclonal antibodies in research?

Purified PTPRC monoclonal antibodies are extensively used in several research applications:

• Flow cytometry for immunophenotyping of leukocyte populations

• Immunohistochemistry of tissue sections for detecting immune cell infiltration

• Western blotting for protein expression analysis

• Immunoprecipitation studies for protein-protein interaction analysis

• Functional studies examining phosphatase activity

• Cell sorting and enrichment of specific immune cell populations

• Tracking immune responses in experimental disease models

The clone HI30 (mentioned in search results) is particularly useful for flow cytometry applications, allowing researchers to identify and characterize CD45-positive populations in various experimental settings .

How should PTPRC monoclonal antibodies be stored to maintain optimal activity?

For optimal stability and activity retention:

• Store purified PTPRC monoclonal antibodies at -20°C for long-term storage

• For conjugated antibodies (e.g., APC-conjugated), store at 4°C protected from light

• Avoid repeated freeze-thaw cycles by aliquoting the antibody upon first thaw

• Store working dilutions at 4°C for no more than one week

• Follow manufacturer's specific recommendations for each clone and formulation

• Monitor for signs of degradation (reduced signal intensity, increased background)

How should I design a flow cytometry panel incorporating PTPRC/CD45 antibodies?

When designing a flow cytometry panel with PTPRC/CD45 antibodies:

Panel Design Strategy:

• Consider CD45 as a core marker for identifying all leukocytes

• Select appropriate fluorochromes based on your cytometer configuration and other markers

• For multicolor panels, place CD45 on a bright fluorochrome if analyzing rare populations

• Include appropriate compensation controls

• Consider potential spectral overlap with other markers in your panel

Gating Strategy Example:

This approach enables robust identification of leukocyte populations for downstream analysis of specific cell subsets .

What controls should I include when using PTPRC antibodies for flow cytometry?

To ensure reliable and interpretable results:

Essential Controls:

• Isotype control matched to the PTPRC antibody's isotype, species, and fluorochrome

• Fluorescence Minus One (FMO) control (all antibodies except anti-PTPRC)

• Single-stained compensation controls for each fluorochrome

• Unstained control for autofluorescence assessment

• Positive control (sample known to express PTPRC)

• Negative control (sample or cell line not expressing PTPRC)

Validation Controls:

• Titration experiment to determine optimal antibody concentration

• Blocking experiment to confirm specificity (pre-incubate with unlabeled antibody)

• Cross-reactivity assessment if working with non-human species

These controls help distinguish specific staining from background and ensure accurate data interpretation .

How can I optimize immunohistochemistry protocols using PTPRC monoclonal antibodies?

For optimal IHC results with PTPRC monoclonal antibodies:

Protocol Optimization Steps:

• Antigen Retrieval:

  • Test both heat-induced (citrate buffer pH 6.0 or EDTA pH 9.0) and enzymatic methods

  • Optimize retrieval time (typically 15-30 minutes)

• Antibody Dilution:

  • Test serial dilutions (typically 1:50 to 1:500) to determine optimal concentration

  • Incubate overnight at 4°C for maximum sensitivity

• Detection Systems:

  • For low expression, use polymer-based or amplification systems

  • For quantitative analysis, use chromogenic substrates with consistent development times

• Controls:

  • Include lymphoid tissue as positive control

  • Use non-immune IgG of same species and concentration as negative control

• Counterstain Optimization:

  • Adjust hematoxylin intensity to allow clear visualization of membrane staining

This systematic approach helps achieve consistent and specific staining for PTPRC in tissue sections.

What are common issues when using PTPRC monoclonal antibodies in flow cytometry and how can they be resolved?

Researchers commonly encounter these issues:

Problem: Low or No Signal

• Possible causes: Antibody degradation, insufficient permeabilization, epitope masking

• Solutions:

  • Verify antibody activity with positive control

  • Optimize permeabilization if detecting intracellular epitopes

  • Try alternative clone recognizing different epitope

  • Increase antibody concentration within recommended range

Problem: High Background

• Possible causes: Non-specific binding, Fc receptor binding, dead cells

• Solutions:

  • Include Fc receptor blocking step

  • Add viability dye to exclude dead cells

  • Reduce antibody concentration

  • Modify washing steps (increase number or volume)

Problem: Inconsistent Results Between Experiments

• Possible causes: Variations in sample processing, antibody degradation, instrument variability

• Solutions:

  • Standardize sample preparation protocols

  • Include calibration beads for instrument standardization

  • Use internal controls for normalization

  • Maintain antibody storage conditions

These troubleshooting approaches help resolve common technical issues and improve data quality and reproducibility.

How can I distinguish between different isoforms of PTPRC using monoclonal antibodies?

PTPRC exists in multiple isoforms (CD45RA, CD45RB, CD45RC, CD45RO) due to alternative splicing:

Isoform Discrimination Strategy:

Experimental Approaches:

• Two-color flow cytometry with pan-CD45 and isoform-specific antibodies

• Western blot analysis to detect molecular weight differences between isoforms

• RT-PCR to confirm isoform expression at mRNA level

Data Interpretation:

• Consider expression patterns in context of cell activation state

• Use known positive controls for each isoform

• Validate with complementary techniques (e.g., flow + PCR)

This approach enables researchers to monitor immune cell differentiation states through isoform expression patterns.

What considerations are important when using PTPRC antibodies for studying phosphatase activity?

When investigating PTPRC/CD45 phosphatase activity:

Experimental Considerations:

• Antibody Selection:

  • Choose clones that don't interfere with the phosphatase domain

  • Consider using antibodies that modulate phosphatase activity for functional studies

• Activity Assays:

  • Use phosphatase assays with specific substrates

  • Monitor tyrosine phosphorylation status of known PTPRC substrates (Lck, Fyn)

  • Combine with immunoprecipitation to isolate PTPRC before activity assessment

• Controls:

  • Include phosphatase inhibitors as negative controls

  • Use recombinant PTPRC domains as positive controls

  • Consider PTPRC knockout or knockdown systems for specificity validation

• Data Analysis:

  • Normalize phosphatase activity to PTPRC expression level

  • Account for potential contributions from other phosphatases

  • Consider kinetic measurements rather than endpoint assays

This methodological approach allows for meaningful investigation of PTPRC's enzymatic function in various experimental contexts.

How can PTPRC monoclonal antibodies be used in single-cell analysis technologies?

PTPRC antibodies can be integrated into cutting-edge single-cell technologies:

Single-Cell Applications:

• Mass Cytometry (CyTOF):

  • Metal-conjugated anti-PTPRC antibodies enable high-parameter analysis

  • Combine with 30+ other markers for comprehensive immune profiling

  • Use as a lineage marker in deep immunophenotyping panels

• Single-Cell RNA-Seq with Protein Detection:

  • CITE-seq applications using oligonucleotide-tagged PTPRC antibodies

  • Correlate protein expression with transcriptome at single-cell resolution

  • Monitor PTPRC isoform switching events with complementary RNA data

• Imaging Mass Cytometry:

  • Spatial distribution of PTPRC+ cells within tissue microenvironment

  • Multiplex with tumor markers to study immune infiltration patterns

  • Quantify distances between PTPRC+ cells and other cell types

Technical Considerations:

• Antibody clone selection based on epitope accessibility in fixed cells

• Titration optimization for each platform

• Batch effect monitoring with appropriate controls

These applications allow researchers to study PTPRC in heterogeneous cell populations with unprecedented resolution.

What are the considerations for using PTPRC antibodies in studying rare cell populations?

For detecting and analyzing rare PTPRC-expressing populations:

Methodological Approaches:

• Pre-enrichment Strategies:

  • Magnetic separation using PTPRC antibodies prior to analysis

  • Density gradient separation of leukocytes from non-hematopoietic cells

  • Depletion of abundant populations to enrich rare subsets

• High-Dimensional Flow Cytometry:

  • Panel design with PTPRC on bright fluorochromes

  • Include additional markers for fine discrimination of rare subsets

  • Acquire sufficient events (≥1 million) to capture rare populations

• Sensitivity Enhancement:

  • Signal amplification systems for dim PTPRC expression

  • Multi-laser excitation for improved signal resolution

  • Optimization of PMT voltages for maximum dynamic range

Analysis Considerations:

• Use specialized algorithms (viSNE, SPADE, FlowSOM) for unbiased detection

• Validate rare populations with orthogonal techniques

• Calculate absolute cell counts using counting beads

This systematic approach enables reliable identification and characterization of rare PTPRC-expressing cell populations in complex samples.

How can PTPRC antibodies be used to study the tumor immune microenvironment?

PTPRC antibodies are valuable tools for tumor immunology research:

Research Applications:

• Immune Infiltrate Characterization:

  • Quantify total leukocyte infiltration using PTPRC as a pan-leukocyte marker

  • Combine with lineage markers to determine composition of immune infiltrate

  • Assess spatial distribution of immune cells within tumor regions

• Functional Studies:

  • Monitor activation status through PTPRC isoform expression

  • Correlate PTPRC+ cell density with treatment response

  • Isolate tumor-infiltrating leukocytes for ex vivo functional assays

• Methodology Integration:

  • Multiplex immunohistochemistry with PTPRC and tumor markers

  • Flow cytometry of disaggregated tumor tissue

  • Single-cell analysis of PTPRC+ cells from the tumor microenvironment

Data Interpretation Framework:

This comprehensive approach provides insights into immune-tumor interactions and potential therapeutic targets.

How can PTPRC antibodies contribute to COVID-19 and infectious disease research?

PTPRC monoclonal antibodies offer valuable tools for infectious disease research:

COVID-19 Applications:

• Immune Monitoring:

  • Track dynamics of leukocyte populations during infection

  • Identify immune signatures associated with disease severity

  • Monitor immune reconstitution during recovery

• Mechanistic Studies:

  • Investigate PTPRC isoform switching in response to SARS-CoV-2

  • Study phosphatase activity modulation in infected immune cells

  • Examine immune exhaustion phenotypes using PTPRC isoform patterns

• Methodological Approaches:

  • Whole blood immunophenotyping with minimal manipulation

  • Longitudinal studies of immune subsets during disease progression

  • Integration with cytokine measurement for comprehensive immune profiling

This research direction can yield insights into pathogen-host interactions and inform therapeutic development.

What are recent developments in PTPRC antibody applications for hematopoietic stem cell research?

Recent innovations in stem cell research using PTPRC antibodies include:

Advanced Applications:

• HSC Identification:

  • Precise delineation of HSC populations using PTPRC with stem cell markers

  • Differential PTPRC expression levels to distinguish primitive vs. committed progenitors

  • PTPRC isoform expression as indicator of differentiation potential

• HSC Purification:

  • Negative selection strategies using PTPRC for mesenchymal stem cell isolation

  • Positive selection of defined hematopoietic progenitor populations

  • Combined positive/negative selection approaches for highly purified populations

• Transplantation Studies:

  • Tracking donor vs. recipient cells using PTPRC allelic variants (CD45.1/CD45.2)

  • Monitoring engraftment kinetics of specific hematopoietic lineages

  • Assessment of chimerism in xenograft models

Methodological Framework:

These approaches advance our understanding of normal and malignant hematopoiesis.

How might PTPRC antibodies be utilized in neuroinflammation and neuroimmunology research?

PTPRC antibodies have significant utility in neuroinflammation research:

Neuroimmunology Applications:

• CNS Immune Cell Identification:

  • Distinguish resident microglia (PTPRC^low) from infiltrating leukocytes (PTPRC^high)

  • Characterize infiltrating immune populations in neuroinflammatory disorders

  • Monitor blood-brain barrier integrity through tracking PTPRC+ cell infiltration

• Disease Models:

  • Track immune infiltration dynamics in multiple sclerosis models

  • Characterize neuroinflammatory responses in neurodegenerative diseases

  • Evaluate therapeutic efficacy in reducing CNS immune infiltration

• Methodological Approaches:

  • Flow cytometry of isolated CNS immune cells

  • Multiplex immunohistochemistry for spatial context

  • Live imaging of PTPRC+ cell trafficking in experimental models

Experimental Design Considerations:

• Tissue preparation methods that preserve surface epitopes

• Appropriate controls distinguishing resident vs. infiltrating cells

• Integration with markers of activation and tissue damage

This research area provides insights into neurological disease mechanisms and potential therapeutic targets for neuroinflammatory conditions.