Normal human peripheral blood cells were unstimulated (left) or stimulated (right) with immobilized Anti-Human CD3 Functional Grade Purified (Product # 16-0037), soluble Anti-Human CD28 Functional Grade Purified (Product # 16-0289), and Human IL-2 Recombinant Protein (Product # 14-8029) for 24 hours. Cells were surface stained with Anti-Human CD4 APC (Product # 17-0049) and Anti-Human GARP FITC. Cells were then fixed and permeabilized with the Foxp3/Transcription Factor Staining Buffers (Product # 00-5523) followed by intracellular staining with Anti-Human Foxp3 eFluor® 450 (Product # 48-4776). CD4+ cells in the lymphocyte gate were used for analysis.
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rat / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||5 µL (0.5 µg)/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Description: The G14D9 monoclonal antibody reacts with human Glycoprotein A Repetitions Predominant (GARP, also known as LRRC32 or Garpin). GARP is an approximately 80 kDa glycoprotein that is expressed on the cell surface. Using northern blot, RT-PCR or microarray analyses, the expression of GARP has been reported in placenta, lung, kidney, heart, ovary, liver, skeletal muscle, and pancreas. Protein expression has been observed on megakaryocytes, platelets and activated regulatory T (Treg) cells. The expression of GARP on the surface of activated Treg cells has been reported to be necessary for their suppressive function, possibly related to its role as a cell surface receptor for LAP/TGF beta.
Applications Reported: This G14D9 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This G14D9 antibody has been pre-titrated and tested by flow cytometric analysis of stimulated human peripheral blood cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser
GARP, also known as leucine rich repeat containing 32, is a type I membrane protein with twenty-two leucine-rich repeats and a short intracytoplasmic region. GARP was identified based on the analysis of gene expression profiling of Treg and Th cells following TCR stimulation. Since GARP is expressed in CD4+, CD25+, FoxP3+ cells (Tregs), it may be a candidate for cell surface marker for Treg cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
||Phenotypic and functional characteristics of CD4+ CD39+ FOXP3+ and CD4+ CD39+ FOXP3neg T-cell subsets in cancer patients.||Schuler PJ,Schilling B,Harasymczuk M,Hoffmann TK,Johnson J,Lang S,Whiteside TL||European journal of immunology (42:1876)||2012|
|Not Applicable||Not Cited||Phenotypic and functional characteristics of CD4+ CD39+ FOXP3+ and CD4+ CD39+ FOXP3neg T-cell subsets in cancer patients.||Schuler PJ,Schilling B,Harasymczuk M,Hoffmann TK,Johnson J,Lang S,Whiteside TL||European journal of immunology (42:1876)||2012|