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Detection of human IFN-gamma and TNF-a cytokine-producing human CD4 T cells using the Attune® Acoustic Focusing Cytometer. Red blood cell lysed peripheral blood mononuclear cells were left unstimulated (left column) or stimulated (right column) for 5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. Cells were then fixed and permeabilized using the FIX & PERM® Cell Permeabilization Kit (Cat. no. GAS003 or GAS004) and stained intracellularly with anti-human CD4 APC (Cat. no. MHCD0405) and either anti-human interferon gamma (IFN-gamma) FITC (Cat. no. MHCIFG01) (A) or anti-human tumor necrosis factor alpha (TNF-a) FITC (Cat. no. A18469) (B). Samples were collected using the Attune® Acoustic Focusing Cytometer (blue/red) with 488 nm excitation and 530/30 nm bandpass emission filter to detect FITC. 633 nm excitation and 660/20 nm bandpass emission filter were used to detect APC.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Storage buffer||PBS with BSA, sucrose|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay-Dependent|
* 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.
Allophycocyanin (APC) is a stable and highly soluble phycobiliprotein that provides maximal absorbance and fluorescence without susceptibility to internal or external fluorescence quenching, thus providing exceptional quantum yields and molar extinction coefficients.
The CD4 antigen is involved in the recognition of the type II MHC antigen. It is also a receptor for HIV. It is present on most T helper cells and normal thymocytes. The cytoplasmic portion of CD4 is associated with p56lck tyrosine kinase. CD4 expression is commonly found in human lymph nodes and tonsils. CD44 is a type 1 transmembrane glycoprotein also known as Phagocytic Glycoprotein-1 (pgp-1) and HCAM. CD44 is the receptor for hyaluronate and exists as a large number of different isoforms due to alternative RNA splicing. The major isoform expressed on lymphocytes, myeloid cells, and erythrocytes is a glycosylated type 1 transmembrane protein. Other isoforms contain glycosaminoglycans and are expressed on hematopoietic and non-hematopoietic cells. CD44 is involved in adhesion of leukocytes to endothelial cells, stromal cells, and the extracellular matrix.
Analyte Specific Reagent
Nef neutralizes the ability of exosomes from CD4+ T cells to act as decoys during HIV-1 infection.
MHCD0405 was used in flow cytometry and immunocytochemistry to test if Nef modifies the composition of exosomes released by T lymphocytes
|de Carvalho JV,de Castro RO,da Silva EZ,Silveira PP,da Silva-Januário ME,Arruda E,Jamur MC,Oliver C,Aguiar RS,daSilva LL||PloS one (9:null)||2014|
An HIV-encoded antisense long noncoding RNA epigenetically regulates viral transcription.
MHCD0405 was used in flow cytometry to characterize the effects of human immunodeficiency virus lncRNA expression
|Saayman S,Ackley A,Turner AM,Famiglietti M,Bosque A,Clemson M,Planelles V,Morris KV||Molecular therapy : the journal of the American Society of Gene Therapy (22:1164)||2014|
Human tetherin exerts strong selection pressure on the HIV-1 group N Vpu protein.
MHCD0405 was used in flow cytometry to examine the function of Vpu proteins from seven HIV group N strains.
|Sauter D,Unterweger D,Vogl M,Usmani SM,Heigele A,Kluge SF,Hermkes E,Moll M,Barker E,Peeters M,Learn GH,Bibollet-Ruche F,Fritz JV,Fackler OT,Hahn BH,Kirchhoff F||PLoS pathogens (8:null)||2012|
Characterization of an HIV-targeted transcriptional gene-silencing RNA in primary cells.
MHCD0405 was used in flow cytometry to demonstrate that transcriptional gene silencing reduces HIV transcription in primary human CD4(+) T cells.
|Turner AM,Ackley AM,Matrone MA,Morris KV||Human gene therapy (23:473)||2012|
Reduced cell surface expression of CCR5 in CCR5Delta 32 heterozygotes is mediated by gene dosage, rather than by receptor sequestration.
MHCD0405 was used in flow cytometry to determine how CCR5 expression is reduced in cells from ccr5Delta32 heterozygotes.
|Venkatesan S,Petrovic A,Van Ryk DI,Locati M,Weissman D,Murphy PM||The Journal of biological chemistry (277:2287)||2002|
A membrane-proximal basic domain and cysteine cluster in the C-terminal tail of CCR5 constitute a bipartite motif critical for cell surface expression.
MHCD0405 was used in flow cytometry to characterize CCR5 truncations.
|Venkatesan S,Petrovic A,Locati M,Kim YO,Weissman D,Murphy PM||The Journal of biological chemistry (276:40133)||2001|
CD4 antigen (p55); CD4 antigen p55; CD4 receptor; T-cell surface antigen T4/Leu-3