- 50-0577-42 - Invitrogen Antibodies B3GAT1 antibody

Submitted references Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir.

Neidleman J, Luo X, Frouard J, Xie G, Hsiao F, Ma T, Morcilla V, Lee A, Telwatte S, Thomas R, Tamaki W, Wheeler B, Hoh R, Somsouk M, Vohra P, Milush J, James KS, Archin NM, Hunt PW, Deeks SG, Yukl SA, Palmer S, Greene WC, Roan NR
eLife 2020 Sep 29;9

Mass Cytometric Analysis of HIV Entry, Replication, and Remodeling in Tissue CD4+ T Cells.

Cavrois M, Banerjee T, Mukherjee G, Raman N, Hussien R, Rodriguez BA, Vasquez J, Spitzer MH, Lazarus NH, Jones JJ, Ochsenbauer C, McCune JM, Butcher EC, Arvin AM, Sen N, Greene WC, Roan NR
Cell reports 2017 Jul 25;20(4):984-998

A novel method for autophagy detection in primary cells: impaired levels of macroautophagy in immunosenescent T cells.

Phadwal K, Alegre-Abarrategui J, Watson AS, Pike L, Anbalagan S, Hammond EM, Wade-Martins R, McMichael A, Klenerman P, Simon AK
Autophagy 2012 Apr;8(4):677-89

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Supportive validation

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Experimental details: Staining of normal human peripheral blood cells with Anti-Human CD4 PE-Cyanine7 (Product # 25-0047-42) and Mouse IgM Isotype Control eFluor® 660 (Product # 50-4752-82) (left) or Anti-Human CD57 eFluor® 660 (right). Cells in the lymphocyte gate were used for analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Design of tailored sort strategies to isolate three populations memory CD4+ PD1+ T cells harboring different frequencies of kNN latent cells. Sort strategies were designed for each of the four leukapheresis donors analyzed by PP-SLIDE. Shown are the CyTOF datasets, the unstimulated atlas cells are shown as gray contours and kNN latent cells shown as red contours. Gates in black correspond to upstream gates. Each sorting strategy isolates three populations of memory CD4+ PD1+ T cells: two disenriched ( pink , purple ), and one enriched ( red ). Results were pre-gated on live, singlet memory CD4+ T cells (CD3+CD8-CD19-CD45RO+CD45RA-). The three functional assays applied to sorted cells are listed. The gray inset on the right shows frequencies of kNN latent cells in the final enriched populations (per million memory CD4+ T cells) and the fold-enrichment of kNN latent cells in the final sorted enriched population relative to each of the disenriched populations. In instances where a disenriched population did not harbor any kNN cells, the fold-enrichment is listed as NA (not available) because the fold-enrichment is infinity when divided by zero.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5. Aging and replicative senescence markers in human T lymphocytes. PBMCs from healthy young (< 28 y) and old (> 56 y) donors were stained for CD28 and CD57 and run on LSR II flow cytometer. (A) Autophagy levels (Mean BDS) in CD8 + T cells from young (< 28 y, n = 8) and old (> 56 y, n = 8) donors under basal and basal+I (for 2 h) conditions (mean +- SEM, *p < 0.0499 between young and old basal+I). (B) Representative dot plots from a young and an old donor showing percentages of CD28 and CD57 cells gated on CD8 + T cells. (C) Bar graph showing % of CD8 + lymphocytes with CD28 and CD57 markers in four young and old donors (mean +- SEM, p = 0.0571 for CD8 CD28 population and *p = 0.0286 for CD8 CD57 population). (D) Overlaid histogram of gammaH2AX (DNA double-strand break) levels of CD8 + lymphocytes from three young and old donors gated on CD28 + CD57 - population (geometric mean +- SEM, *p = 0.0286). (E) Overlaid histogram of FAS (CD95) levels of CD8 + lymphocytes from four young and old donors gated on CD28 + CD57 - population (geometric mean +- SEM, *p = 0.0286). PBMCs from four healthy young and old donors were cultured under control and starved conditions for 2 h and stained for CD8, CD57, LC3 and Lyso-ID. (F) Colocalization of LC3 and lysosomal marker in CD8 + CD57 +/- cells, expressed as mean BDS ratio between starved and basal treatments (mean +- SEM, n = 5 (young donors), n = 8 (old donors), **p = 0.0049, *p = 0.035).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6. Levels of gammaH2AX foci in senescent T cells with low autophagy. PBMCs from three healthy donors were bead sorted for CD8 + T cells and stained for CD57 gammaH2AX, LC3 and Lyso-ID and run on ImageStream. (A) Representative BDS overlay histogram for CD8 + CD57 + (red) for CD8 + CD57 - (green) depicts the low and high autophagy gates. (B) Bar graph showing proportion of CD8 + CD57 +/- lymphocytes with low and high autophagy in three healthy donors (mean +- SEM, **p = 0.0029, *p = 0.0273). (C) Bar graph showing mean gammaH2AX foci/cell in low and high autophagy CD8 + CD57 + , CD8 + CD57 - populations (geometric mean +- SEM, **p = 0.0057 and **p = 0.0087, respectively). (D) Representative ImageStream images of gammaH2AX foci in CD8 + lymphocytes from young and old cells.

50-0577-42

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