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 and 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||Non-human primate, 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|
Deletion of cIAP1 and cIAP2 in murine B lymphocytes constitutively activates cell survival pathways and inactivates the germinal center response.
MHCD0405 was used in flow cytometry to determine the role of cIAP1 and cIAP2 to B-cell physiology in vivo.
|Gardam S,Turner VM,Anderton H,Limaye S,Basten A,Koentgen F,Vaux DL,Silke J,Brink R||Blood (117:4041)||2011|
|Non-human primate||Not Cited||
Subcutaneous versus intravenous administration of rituximab: pharmacokinetics, CD20 target coverage and B-cell depletion in cynomolgus monkeys.
MHCD0405 was used in flow cytometry to assess subcutaneous administration of rituximab.
|Mao CP,Brovarney MR,Dabbagh K,Birnböck HF,Richter WF,Del Nagro CJ||PloS one (8:null)||2013|
Human neonatal naive CD4+ T cells have enhanced activation-dependent signaling regulated by the microRNA miR-181a.
MHCD0405 was used in flow cytometry to test if impaired human neonatal CD4(+) T cell immunity is due to reduced signaling by naive CD4(+) T cells following engagement of the αβ-TCR/CD3 complex and CD28.
|Palin AC,Ramachandran V,Acharya S,Lewis DB||Journal of immunology (Baltimore, Md. : 1950) (190:2682)||2013|
Helicobacter pylori induces in-vivo expansion of human regulatory T cells through stimulating interleukin-1ß production by dendritic cells.
MHCD0405 was used in flow cytometry to elucidate the influence of H. pylori on T(reg) function and proliferation.
|Mitchell PJ,Afzali B,Fazekasova H,Chen D,Ali N,Powell N,Lord GM,Lechler RI,Lombardi G||Clinical and experimental immunology (170:300)||2012|
Phenotypic and molecular characterization of CD103+ CD4+ T cells in bronchoalveolar lavage from patients with interstitial lung diseases.
MHCD0405 was used in flow cytometry to test if CD103 has a regulatory role in the microenvironment of the epithelial cell layer.
|Braun RK,Foerster M,Grahmann PR,Haefner D,Workalemahu G,Kroegel C||Cytometry. Part B, Clinical cytometry (54:19)||2003|
Graft-versus-leukemia effect after suicide-gene-mediated control of graft-versus-host disease.
MHCD0405 was used in flow cytometry to investigate graft-versus-leukemia.
|Litvinova E,Maury S,Boyer O,Bruel S,Benard L,Boisserie G,Klatzmann D,Cohen JL||Blood (100:2020)||2002|
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|
Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages.
MHCD0405 was used in flow cytometry to determine the number of CD4, CCR5, and CXCR4 antibody-binding sites on various T cells and macrophages.
|Lee B,Sharron M,Montaner LJ,Weissman D,Doms RW||Proceedings of the National Academy of Sciences of the United States of America (96:5215)||1999|