50-5273-41

antibody from Invitrogen Antibodies

Targeting: CD8B CD8B1

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

50-5273-41

Provider

Invitrogen Antibodies

Product name

CD8b Monoclonal Antibody (SIDI8BEE), eFluor™ 660, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The SIDI8BEE monoclonal antibody reacts with human and rhesus macaque CD8 beta. The CD8b chain associates with the CD8a chain to form the CD8ab heterodimer expressed on the surface of a majority of thymocytes, and on peripheral cytotoxic alpha beta TCR T cells. The CD8 receptor complex binds to MHC class I and plays a role in T cell development and activation of mature T cells. Long been thought to exist either as a heterodimer of the alpha and beta chain or as a homodimer of alpha chains, beta chains have now been shown to homodimerize as well. SIDI8BEE can detect the CD8 beta chain in both the CD8ab heterodimer and the CD8bb homodimer.

Antibody clone number

SIDI8BEE

Concentration

5 µL/Test

References

Submitted references

Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities.

Van Caeneghem Y, De Munter S, Tieppo P, Goetgeluk G, Weening K, Verstichel G, Bonte S, Taghon T, Leclercq G, Kerre T, Debets R, Vermijlen D, Abken H, Vandekerckhove B
Oncoimmunology 2017;6(3):e1283460

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GATA3 induces human T-cell commitment by restraining Notch activity and repressing NK-cell fate.

Van de Walle I, Dolens AC, Durinck K, De Mulder K, Van Loocke W, Damle S, Waegemans E, De Medts J, Velghe I, De Smedt M, Vandekerckhove B, Kerre T, Plum J, Leclercq G, Rothenberg EV, Van Vlierberghe P, Speleman F, Taghon T
Nature communications 2016 Apr 6;7:11171

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of normal human peripheral blood cells either with Anti-Human CD4 FITC (Product # 11-0048-42) (left) or Anti-Human CD8a (Product # 12-0086-42) (right) and Anti-Human/Non-Human Primate CD8b eFluor® 660. Cells in the lymphocyte gate were used for analysis.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 GATA3 restrains Notch activation at the T-lineage commitment stage. ( a ) Quantitative PCR of GATA3 , DTX1 , HES1 and TCF7 expression in different stages of in vitro generated T cell precursors from CB CD34 + lin - HPCs after 7 days of OP9-DLL4 coculture. Data shows the average expression in 3-4 independent samples on a log scale and erros bars indicate s.e.m. ( b , c ) GSEA shows a significant enrichment of the top 500 Notch-dependent genes in human CD34 + thymocytes in the set of genes higher expressed in ( b ) uncommitted CD34 + CD1a - versus CD34 + CD1a + committed T-cell precursors 50 , and genes expressed higher in ( c ) control versus GATA3-transduced CD34 + thymocytes as determined by microarray after 48 h of transduction. ( d ) Flow cytometry analysis of control and GATA3 -transduced CD34 + CD1 - uncommitted thymocytes in OP9-DLL1 cocultures with addition of 0 or 1 muM GSI and in the presence of IL7, SCF and FLT3L, showing the development of CD4 + CD8beta + DP thymocytes after 6 days of coculture. ( e ) Graph show absolute number of CD4 + CD8beta + DP thymocytes, generated in corresponding cultures shown in d . Data shows the average of four independent experiments and errors bars show s.e.m. * P

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3. Phenotype and endogenous TCR expression of CD34 + HPC-derived transgenic AR + T cells. Flow cytometric analysis of the AR-transgenic T cells. (A) CAR-transgenic GFP + cells of cultures transduced to express either the CAR:zeta or the CAR:28zeta were analyzed on day 26 of OP9-DL1 culture for CD3 and TCRalphabeta expression. As a control, GFP - cells are shown from the OP9-DL1 culture transduced to express the CAR:zeta ( N = 5). (B) Dot plots show CD3 expression of cells from the OP9-DL1 cultures transgenic for the wtTCR, TCR:zeta and TCR:28zeta. Vbeta14 staining is used to mark transgene expression, as no GFP is expressed by the transgenic cells ( N = 5). (C) Surface and cytoplasmic staining for CD3 of in vitro generated mature T cells that were expanded for one cycle on feeder cells in the presence of cytokines. (D) Expression of various membrane markers by the CD27 + CD1a - mature T cells at the end of OP9-DL1 culture (46 d) ( N = 2). (E) Day 0: fresh cord blood after MACS CD34 enrichment sorted using the sorting window shown. Day 13: cord blood cells cultured on OP9-DL1 were sorted for CD5 CD7 double positive cells, using the indicated sorting window. The cells were then transduced to express CAR:28zeta and further differentiated on OP9-DL1 feeder layer. Day 21: analysis of the transgenic GFP + cultured cells for DP cells and CD27 + CD1a - mature cells. (F) Flow cytometric analysis of GFP + CAR:28zeta-transgenic cultures, gated on GFP + CD27 + CD1a