Anti-CD68 antibody [SP251]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [SP251] (AB192847)

10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section). Merged staining of anti-FOXP3 (ab215206; Cyan; TG540N), anti-PD1 (ab52587; Red; TG700N), anti-CD163 (ab182422; Brown; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Violet; TG620N), anti-CD8 alpha (ab101500; Purple; TG540S), anti-CD20 (ab9475; Grey; TG660S), anti-CD68 (ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain. The inset image shows the separate CD68 signal. The section was incubated in nine rounds of staining; in the order of ab215206 (1/100 dilution), ab52587 (1/200 dilution), ab182422 (1/300 dilution), ab92511 (1/200 dilution), ab133616 (1/600 dilution), ab101500 (1/300 dilution), ab9475 (1/100 dilution), ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system. Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [SP251] (AB192847)

Chromogenic multiplex immunohistochemical staining of FFPE normal human tonsil tissue. ab16667 anti-Ki67 DAB chromogen. ab16669 anti-CD3 epsilon purple chromogen and ab192847 anti-CD68 teal chromogen plus haematoxylin II counterstain. Chromogenic immunostaining was performed on a Roche Ventana Benchmark Ultra instrument. The section was deparaffinised and incubated with CC1 solution for 24min 100°C. Following this with 3 rounds of staining in the order of ab16667 (1/500 dilution), ab192847 (1/4000 dilution) and ab16669 (1/1000 dilution). Between rounds of staining antibody denaturation was conducted using Ultra CC2 solution for 8min at 100oC to avoid cross reactivity. Signal was developed with anti-rabbit HQ followed by anti-HQ HRP coupled with Chromomap DAB kit Discovery purple or Discovery teal chromogens and haematoxylin II counterstain.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD68 antibody [SP251] (AB192847)

Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cells labeling CD68 with ab192847 at 1/100 dilution (green) compared with a negative control rabbit IgG (blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [SP251] (AB192847)

Immunohistochemical analysis of formalin fixed paraffin (FFPE) embedded tonsil labelling CD68 with ab192847 at a dilution of 1/600. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an ChromoMap DAB (RUO) IHC Detection Kit with anti rabbit HQ and anti HQ HRP. Heat mediated antigen retrieval was conducted for 24 min with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5. ab192847 was incubated at 37°C for 16 min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [SP251] (AB192847)

Formalin-fixed paraffin-embedded human tonsil tissue stained for CD68 using ab192847 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [SP251] (AB192847)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD68 with ab192847 at 1/100 dilution (0.184 μg/ml) (incubated for 10 minutes at room temperature). Heat mediated antigen retrieval was performed with sodium citrate buffer (pH 6.0 epitope retrieval solution 1) for 20 minutes. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Negative control : PBS instead of the primary antibody.

Positive staining on the macrophages in human tonsil performed on a Leica Biosystems BOND^® RX instrument.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [SP251] (AB192847)

Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

Merged staining of anti-PD1 (ab237728; orange; Opal™520) anti-PDL1 (ab237726; green; Opal™540) anti-CD68 (ab192847; yellow; Opal™570) anti-CD3 epsilon (ab16669; red; Opal™620) anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT Akoya Biosciences^®).

The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (Leica ER1 pH6.0 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round to avoid any cross-reactivity.

DAPI (dark blue) was used as a nuclear counter stain.

Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.