Figure A. APC conjugated Mouse anti Human CD63 and FITC conjugated Mouse IgG2a isotype control . Figure B. APC conjugated Mouse anti Human CD63 and FITC conjugated Mouse anti Human CD14 . All experiments performed on human blood gated on live, single mononuclear cells in the presence of 10% human serum. Data acquired on the ZE5 Cell Analyzer

Figure A. FITC conjugated Mouse anti Human CD31 and Alexa Fluor647 conjugated Mouse IgG2a isotype contol. Figure B. FITC conjugated Mouse anti Human CD31 and Alexa Fluor647 conjugated Mouse CD14 . All experiments performed on human peripheral blood mononuclear cells in the presence of human SeroBlock

Figure A. Alexa Fluor 488 conjugated Mouse anti Human CD9 and RPE conjugated Mouse IgG2a isotype control . Figure B. Alexa Fluor 488 conjugated Mouse anti Human CD9 and RPE conjugated Mouse anti Human CD14 . All experiments performed on red cell lysed human blood gated on myeloid cells. Data acquired on the ZE5 cell analyzer

Sandwich ELISA analysis of Human CD14 binding using Mouse anti CD14 antibody, clone MEM-18 as a capture reagent and biotinylated Mouse anti Human CD14 antibody, clone Tük4 as a detection reagent with purified CD14 as antigen for generation of the standard curve. Detection is by HRP conjugated streptavidin. Microtitre plate is read at O.D. 450 nm on the Microplate Absorbance Reader . Serum samples diluted 1:200 and 1:400 are shown green and red respectively, while plasma samples also diluted 1:200 and 1:400 are blue and orange respectively

Phycoerythrin conjugated Mouse anti Human CD14 antibody, clone Tük4 used for the evaluation of CD14 expression on bovine monocytes by flow cytometry.

Image caption:

Gating strategy utilized for flow cytometric analysis of bovine peripheral blood leukocytes showing representative data from one animal. After gating on viable (propidium-iodide-negative) cells (A), cell doublets were excluded by SSC-A and SSC-H gating (B). Bovine mononuclear cells (MNC) and granulocytes (PMN) were gated based on their forward and side scatter characteristics and their percentages were calculated (C). Three-color immunofluorescence of bovine MNC with mAbs to CD172a, CD14 and CD16 defines three monocyte subsets in peripheral blood. Viable mononuclear cells, based on forward and side scatter characteristics, were gated on CD172a-positive cells (D). Dot plots of CD14 and CD16 expression display classical monocytes (CD14+CD16?, upper left), intermediate monocytes (CD14+CD16+, upper right) and nonclassical monocytes (CD14?CD16+, lower right) (E)


From: Pomeroy B, Sipka A, Hussen J, Eger M, Schukken Y, Schuberth HJ.

Counts of bovine monocyte subsets prior to calving are predictive for postpartum occurrence of mastitis and metritis.

Vet Res. 2017 Feb 21;48(1):13.

This is from an open access article distributed under the terms of the Creative Commons Attribution License.

Mouse anti Human CD14 antibody clone Tük4 used to evaluate CD14 expression on canine monocytes by flow cytometry.

Image caption:

CD11b+CD14?MHCII? cells suppress T cell proliferation. Facs sorted CD11b+CD14?MHCII? cells isolated from a dog with osteosarcoma or healthy PBMCs were co-incubated with mitogen-stimulated CD4+ and CD8+ T cells isolated from a healthy dog for 72 hs. No stimulated cells were used as negative control. Proliferative responses were measured by 3H-thymidine incorporation from experiments performed in triplicate. CPM, counts per minute. Mean ± SEM are shown.


From: Goulart MR, Pluhar GE, Ohlfest JR (2012)

Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer.

PLoS ONE 7(3): e33274.

This is from an open access article distributed under the terms of the Creative Commons Attribution License.

Mouse anti Human CD14 antibody clone Tük4 used to evaluate CD14 expression on canine monocytes by flow cytometry.

Image caption:

Immunophenotyping gating strategy and morphological analysis for MDSC identification in peripheral blood of dogs. PBMCs from healthy dogs and dogs with cancer were stained for the myeloid marker CD11b, monocytic marker CD14 and MHC II. (A) Representative flow cytometric analysis of forward and side scatter and gated CD11b+CD14?MHCII? cells from dogs with advanced or metastatic tumors compared to dogs with early stage non-metastatic tumors and healthy control dogs. Plots are representative of dog with advanced metastatic hemangiosarcoma (top), early stage bladder transitional cell carcinoma (middle) and a healthy dog. (B) FACS sorted CD11b+CD14?MHCII? cells were stained with diff-quick for cell morphology evaluation. A representative example of polymorphonuclear granulocyte morphology of CD11b+CD14?MHCII? cells is shown at 63× magnification.


From: Goulart MR, Pluhar GE, Ohlfest JR (2012)

Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer.

PLoS ONE 7(3): e33274.

This is from an open access article distributed under the terms of the Creative Commons Attribution License.

Mouse anti Human CD14 antibody clone Tük4 used to evaluate CD14 expression on canine monocytes by flow cytometry.

Image caption:

CD11b+CD14+MHCII? cells demonstrate ability to suppressive T cell proliferation. (A) CD11b+CD14+MHCII? cells were sorted from peripheral blood sample of an osteosarcoma dog (B) and co-cultured with healthy dog PBMCs in the presence of mitogen for 72 hs. Non-stimulated PBMCs were used as negative control and PBMCs co-cultured with healthy PMNs were used to control for the effect of adding cells to the assay. Proliferative responses were measured by 3H-thymidine incorporation. CPM, counts per minute. The experiment was performed in triplicate. Mean ± SEM are shown.

Mouse anti Human CD14 antibody, clone Tük4 used to identify bovine monocytes in the skin and subcutaneous tissue of adjuvant injected calves by immunofluorescence.

Image caption:

Cellular recruitment to skin and subcutaneous tissues. (A) HE stained sections of skin with subcutaneous tissue from the side injected with adjuvant and the contralateral side, at 24 h post-injection. Scale bars: 200 ?m. (B) Enlargement of outlined areas in A, as indicated. Scale bars: 20 ?m. (C) Immunofluorescent labeling of subcutaneous tissue on the injected side with antibody against CD14 (green). Scale bar: 20 ?m.


From: Lund H, Boysen P, Ã…kesson CP, Lewandowska-Sabat AM and Storset AK (2016)

Transient Migration of Large Numbers of CD14++ CD16+ Monocytes to the Draining Lymph Node after Onset of Inflammation.

Front. Immunol. 7:322.

This is from an open access article distributed under the terms of the Creative Commons Attribution License.