Mouse Anti-CD151 Recombinant Antibody (clone TS151r) (CAT#: HPAB-0212-CN)

Figure 1 α3 and α6 integrins mask the TS151r epitope.

K562 cells were transiently transfected with α3 integrin or vector alone, and then the presence or absence of α3 was verified by flow cytometry using mAb IIF5. Cells lacking or expressing α3 were also stained using CD151 mAbs 5C11 (5) and TS151r (T) (middle and right, respectively).

Kazarov, A. R., Yang, X., Stipp, C. S., Sehgal, B., & Hemler, M. E. (2002). An extracellular site on tetraspanin CD151 determines α3 and α6 integrin–dependent cellular morphology. The Journal of cell biology, 158(7), 1299-1309.

Figure 2 Cell lysis in Triton X-100 and Brij 96 yields distinct populations of CD151-associated integrins.

HT1080 cells were labeled with [35S]methionine for 2.5, 5, 25, and 56 h. Cells were lysed in 1% Triton X-100 or Brij 96, and then immunoprecipitations of α3, CD151, and CD81 were performed using the indicated antibodies. Samples were resolved on 8-15% SDS-PAGE under reducing conditions and exposed to BioMax MS film. Note that during maturation, integrin α3 and α6 subunits are proteolytically cleaved, but fragments remain disulfide bonded. In reducing conditions, one can observe conversion of the ∼150-kD precursor form to fragments of ∼120 and 30 kD (not depicted). Labeled β1 subunit (110 kD), appearing at later time points, closely comigrates with 120 kD α subunit fragments. As seen elsewhere, CD81 associated bands of ∼120 kD from HT1080 cells (such as in panel e) have been clearly identified as corresponding to α3 and α6, with α3 being ∼2-3-fold more abundant than α6 in HT1080 cells.

Kazarov, A. R., Yang, X., Stipp, C. S., Sehgal, B., & Hemler, M. E. (2002). An extracellular site on tetraspanin CD151 determines α3 and α6 integrin–dependent cellular morphology. The Journal of cell biology, 158(7), 1299-1309.

Figure 3 Immunohistochemical staining of smooth and striated muscle tissue with P48 and TS151R against CD151.

Cryostat sections of human smooth (A,C) and striated muscle (B,D) were stained by immunoperoxidase reaction with P48 (A,C) or TS151R (B,D). Both types of muscle cells were strongly stained by P48. Intense staining of CD151 by TS151R was only seen in smooth muscle cells, no staining is found in skeletal muscle. n, nerve; v, vessel. Bar, 10 μm.

Sterk, L. M., Geuijen, C. A., van den Berg, J. G., Claessen, N., Weening, J. J., & Sonnenberg, A. (2002). Association of the tetraspanin CD151 with the laminin-binding integrinsα 3β1, α6β1, α6β4 and α7β1 in cells in culture and in vivo. Journal of cell science, 115(6), 1161-1173.

Figure 4 Reactivity of CD151 with antibody TS151R is cell type specific.

Cryostat sections of human kidney, skin and heart muscle were processed for double immunofluorescence confocal microscopy using TS151R (A,G,J) and P48 (D) against CD151, GoH3 against α6 (E), 439-9B against β4 (H) and polyclonal antibodies against α3 (B) or α7 (K). The composite images (C,F,I,L) were generated by superimposition of the green and red signals, with areas of overlap appearing as yellow. TS151R against CD151 reacts with CD151 in tubular epithelial cells but hardly reacts in glomerular cells (A-C), keratinocytes (G-I) and cardiomyocytes (J-L). P48 against CD151 is used in the skin for comparison and shows a prominent staining especially at the dermo-epidermal junction (D-F). Bar, 10 μm.

Sterk, L. M., Geuijen, C. A., van den Berg, J. G., Claessen, N., Weening, J. J., & Sonnenberg, A. (2002). Association of the tetraspanin CD151 with the laminin-binding integrinsα 3β1, α6β1, α6β4 and α7β1 in cells in culture and in vivo. Journal of cell science, 115(6), 1161-1173.

Figure 5 Expression of laminin-binding integrins on transfected K562 cells increases CD151 cell surface expression.

Flow cytometric analysis of wild-type K562 cells endogenously expressing α5β1 and transfected K562 also expressing αVβ1, α3β1, α6β1, α6β4,α 7X1β1 or α7X2β1. Solid lines in the left panels indicate staining with anti-integrin antibodies: anti-α5 mAb Sam-1 (K562 cells), anti-αV mAb 13C2 (K562-αV), anti-α3 mAb J143 (K562-α3), anti-α6 mAb GoH3 (K562-α6), anti-β4 mAb 439-9B (K562-α6β4) and anti-α7 mAb CA25 (K562-α7BX1 and K562-α7BX2). Staining of cells with anti-CD151 antibodies is shown in the right panels. Solid lines represent staining of cells with mAb TS151R and broken lines with mAb P48. A negative control (dotted line) with secondary goat anti-mouse or rat IgG alone is shown in each panel. R represents the ratio of the mean fluorescence intensity of cells stained with mAb P48 and with secondary antibody alone. A three- to sixfold increase of CD151 expression is observed in transfected cells that ectopically express laminin-binding integrins. The TS151R epitope is masked in all cells expressing one of the laminin-binding integrins.

Sterk, L. M., Geuijen, C. A., van den Berg, J. G., Claessen, N., Weening, J. J., & Sonnenberg, A. (2002). Association of the tetraspanin CD151 with the laminin-binding integrinsα 3β1, α6β1, α6β4 and α7β1 in cells in culture and in vivo. Journal of cell science, 115(6), 1161-1173.

Figure 6 CD151: integrin complexes regulate neurosphere cell self-renewal, migration, and Akt activation.

Equal numbers of viable cells were cultured in 48-well plates with TS151r antibody (or IgG control) for 14 days to form neurospheres. Neurospheres (> 50 μm diameter) were counted. Neurosphere formation was inhibited by TS151r.

Tilghman, J., Schiapparelli, P., Lal, B., Ying, M., Quinones-Hinojosa, A., Xia, S., & Laterra, J. (2016). Regulation of glioblastoma tumor-propagating cells by the integrin partner tetraspanin CD151. Neoplasia, 18(3), 185-198.