- GTX31217 - GeneTex SIRPA antibody

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Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Published customer image: Mouse anti Bovine CD172a antibody, clone CC149 used for the detection of SIRP1a expressing cells in cultured bovine PBMCs by immunofluorescence. Image caption: CD172a cells bind rESAT-6:CFP-10 and M. bovis BCG. CD172a+ cells (PE-labeled) were isolated by high-speed cell sorting (>98% purity) from rESAT-6:CFP-10-stimulated PBMC 6d cultures; incubated with rESAT-6:CFP-10-FITC or BCG-FITC for 2¡V96 hrs; and evaluated by fluorescence microscopy. (A) rESAT-6:CFP-10-FITC bound to the surface of CD172a+ cells in a focal pattern (24 hr cultures shown). Labeling patterns were similar at 2, 24 (shown in Fig. 3A) and 96 hrs after addition of rESAT-6:CFP-10 (FITC) to cells, except for increased polarization of staining at 96 hrs. (B) Over the 96 hr culture period, PE-staining (red) used for CD172a+ cell sorting persisted (24 hr cultures shown) with a polar distribution, possibly due to capping of antibody bound to CD172a. However, rESAT-6:CFP-10-labeling (green) did not overlap with CD172a-PE labeling (red). (C) M. bovis BCG was detected in association with CD172a+ cells at 2 (not shown) and 24 hrs (green, intact bacteria associated with the cell surface, panel C) after addition of the live bacteria to the CD172a+ cell culture and by 96 hrs, M. bovis BCG was internalized and mostly degraded (D). White bar = 10 £gm. From: Waters WR, Palmer MV, Nonnecke BJ, Thacker TC, Estes DM, et al. (2009) Signal Regulatory Protein a (SIRPa)+ Cells in the Adaptive Response to ESAT-6/CFP-10 Protein of Tuberculous Mycobacteria. PLoS ONE 4(7): e6414 .

Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Staining of bovine peripheral blood lymphocytes with Mouse anti Bovine CD172a followed by Goat anti Mouse IgG:FITC

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Staining of bovine peripheral blood monocytes with Mouse anti Bovine CD172a:RPE-Cy5

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Published customer image: Mouse anti Bovine CD172a antibody, clone CC149 used for the detection of SIRP1a expressing cells in bovine PBMCs by flow cytometry. Image caption: Expansion of CD172a+ cells in response to ESAT-6/CFP-10 stimulation. Isolated PBMC were stained with PKH67 and cultured for 6d with media only, rMPB83, a pool of overlapping ESAT-6 and CFP-10 peptides, or rESAT-6:CFP-10. Data are depicted as: (A) dot plots (CD172a-PE (y-axis) versus PKH67 (x-axis, green fluorescence), (B) histograms generated by Modfit Proliferation Wizard analysis of PKH67 staining intensity (gated on CD172a-PE+ cells within the live gate) and (C) mean (¡ÓSEM) percent CD172a+ cells within PBMC cultures (stimulation indicated in the lower margin) from non-infected (open bars, n = 9, includes non- (n = 3), BCG- (n = 3), and RD1- (n = 3) vaccinates) or M. bovis-infected (closed bars, n = 20) cattle. Responses did not differ between controls, BCG- and RD1- vaccinates; thus, these groups were combined. Gate R2 in panel A highlights the CD172a+, PKH67lo proliferative fraction. For panels A and B, data from a single M. bovis-infected animal are provided that are indicative of a representative response. From: Waters WR, Palmer MV, Nonnecke BJ, Thacker TC, Estes DM, et al. (2009) Signal Regulatory Protein a (SIRPa)+ Cells in the Adaptive Response to ESAT-6/CFP-10 Protein of Tuberculous Mycobacteria. PLoS ONE 4(7): e6414 .

GTX31217

Antibody data