Clone:
REA699
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
Peptidase inhibitor 16, CRISP-9, PSP94-binding protein

Extended validation for PI-16 Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA699
CRCBT-02-001++
WCHRI-1++
Cells were incubated with an excess of purified unconjugated PI-16 (REA699) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for PI-16. Human peripheral blood mononuclear cells (PBMCs) were stained with PI-16 antibodies and with a suitable counterstaining. As a control, PI-16 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+/CD25+/CD127low+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for PI-16. Human peripheral blood mononuclear cells (PBMCs) were stained with PI-16 antibodies and with a suitable counterstaining. As a control, PI-16 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+/CD25+/CD127low+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for PI-16. Human peripheral blood mononuclear cells (PBMCs) were stained with PI-16 antibodies and with a suitable counterstaining. As a control, PI-16 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+/CD25+/CD127low+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for PI-16. Human peripheral blood mononuclear cells (PBMCs) were stained with PI-16 antibodies and with a suitable counterstaining. As a control, PI-16 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+/CD25+/CD127low+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using PI-16 (REA699). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using PI-16 (REA699). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using PI-16 (REA699). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for PI-16 Antibody, anti-human, REAfinity™

Overview

Clone REA699 recognizes the human peptidase inhibitor 16 (PI-16) antigen, also known as CRISP-9 or PSP94-binding protein. PI-16 belongs to the Activity cysteine-rich secretory protein (CRISP) family and is expressed in prostate, testis, ovary, and intestine. It is reported to be an independent prognostic marker for prostate cancer. Additionally, PI-16 is over-expressed by CD4
+
/CD25
+
Treg cells, so it identifies a novel distinct subset of functional memory Treg cells, that are able to migrate to sites of inflammation and regulate the pro-inflammatory response.
Additional information: Clone REA699 displays negligible binding to Fc receptors.

Alternative names

Peptidase inhibitor 16, CRISP-9, PSP94-binding protein

Detailed product information

Technical specifications

CloneREA699
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenPI-16
Alternative names of antigenPeptidase inhibitor 16, CRISP-9, PSP94-binding protein
Molecular mass of antigen [kDa]47
Distribution of antigenother, ovary, prostate
Entrez Gene ID221476
RRIDAB_2653192, AB_2653193, AB_2653194, AB_2653195, AB_2653196, AB_2653197, AB_2653198, AB_2653191

References for PI-16 Antibody, anti-human, REAfinity™

Publications

  1. Reeves, J. R. et al. (2005) Identification, purification and characterization of a novel human blood protein with binding affinity for prostate secretory protein of 94 amino acids. Biochem. J. 385(1): 105-114
  2. Nicholson, I. C. et al. (2012) PI16 is expressed by a subset of human memory Treg with enhanced migration to CCL17 and CCL20. Cell. Immunol. 275(1–2): 12-18

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