Anti-CD68 antibody [EPR20545] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue, labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human tonsil is observed (PMID : 19543531). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation. Panel A : merged staining of anti-CD68 (magenta; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human spleen. Secondary antibody was Opal Polymer HRP Ms + Rb, and couterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Clone EPR20545 (ab227458) has been successfully conjugated by Abcam. This image was generated using Anti-CD68 antibody [EPR20545] (Alexa Fluor® 647). Please refer to ab224029 for protocol details.

IHC image of CD68 staining in a section of formalin-fixed paraffin-embedded human prefrontal cortex tissue.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110^oC for 20 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab224029 at 1/1000 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on Kupffer cells of human liver is observed (PMID : 12118106). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Immunofluorescent analysis of 100% methanol-fixed U937 (human histiocytic lymphoma cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U937 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on THP-1 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This IHC data was generated using the same anti-CD68 antibody clone, EPR20545, in a different buffer formulation (cat# ab213363).

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human cervical carcinoma is observed (PMID : 12118106). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Tissue Microarrays stained for "Anti-CD68 antibody [EPR20545]" using "ab213363"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab213363 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Fluorescence multiplex immunohistochemical analysis of the human duodenum (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-liver FABP (ab240401 gray; Opal™690), anti-CD3 epsilon (ab16669 green; Opal™520) and anti-CD68 (ab213363 red; Opal™570) on human duodenum. Panel B : anti-liver FABP stained on enterocytes. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD68 stained on macrophages. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab240401 (1/8000 dilution), ab16669 (1/150 dilution) and ab213363 (1/500 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human thymus tissue labeling CD3 epsilon with ab16669 at 1/500 dilution, LY75/DEC-205 with ab208649 at 1/15000, and CD68 with ab213363 at 1/500 dilution. Panel A : merged staining of anti-CD68 (magenta; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-LY75/DEC-205 (red; Opal™570) on human thymus. Panel B : anti-CD3 epsilon stained on T cells. Panel C : anti-LY75/DEC-205 stained on thymic cortical epithelium and dendritic cells. Panel D : anti-CD68 stained on macrophages. Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of ab213363, ab16669, and ab208649 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI was used as a nuclear counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation. Panel A : merged staining of anti-CD68 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, and counterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-liver FABP (ab240401 gray; Opal™690), anti-CD3 epsilon (ab16669 green; Opal™520) and anti-CD68 (ab213363 red; Opal™570) on human colon. Panel B : anti-liver FABP stained on enterocytes. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD68 stained on macrophages. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab240401 (1/8000 dilution), ab16669 (1/150 dilution) and ab213363 (1/500 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining CD68 with ab213363 at a 1/500 dilution, ab315802 anti-CD34 used at 1/2000 dilution and ab255296 anti-CD177 used at a 1/2000 dilution.

Panel A : merged staining of anti-CD68 (green; Opal™520), anti-CD34 (magenta; Opal™690) and anti-CD177 (gray; Opal™570) on human placenta.

Panel B : anti-CD68 staining macrophages in human placenta.

Panel C : ant-CD34 staining endothelium in human placenta.

Panel D : ant-CD177 staining neutrophils in human placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab213363, ab315802 and ab255296 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human lung tissue staining SFTPA1 + SFTPA2 with ab320648 at a 1/1000 dilution, ab213363 anti-CD68 used at 1/500 dilution and ab307666 anti-Uteroglobin used at a 1/2000 dilution.

Panel A : merged staining of anti-SFTPA1+SFTPA2 (magenta; Opal^™690), anti-CD68 (green; Opal^™520) and anti-Uteroglobin (gray; Opal^™570) on human lung.
Panel B : anti-SFTPA1+SFTPA2 staining alveolar type II cells in human lung.
Panel C : ant-CD68 staining macrophages in human lung.
Panel D : ant-Uteroglobin staining club cells in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab320648, ab211363 and ab307666 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins..

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using the same antibody clone in a different buffer formulation (ab213363).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver cancer tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human liver cancer.
Panel B : anti-CD5L staining Kupffer cells in human liver cancer.
Panel C : anti-CD68 staining Kupffer cells in human liver cancer.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using the same antibody clone in a different buffer formulation (ab213363).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human spleen.
Panel B : anti-CD5L staining Kupffer cells in human spleen.
Panel C : anti-CD68 staining Kupffer cells in human spleen.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213363).

Immunohistochemical analysis of formalin fixed paraffin embedded Human liver labelling active CD68 with ab213363 at a concentration of 0.08 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab213363 Anti CD68 antibody EPR20545 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using the same antibody clone in a different buffer formulation (ab213363).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human liver.
Panel B : anti-CD5L staining Kupffer cells in human liver.
Panel C : anti-CD68 staining Kupffer cells in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] - BSA and Azide free (AB227458)

This data was developed using ab213363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue staining PD-L1 with ab205921 at a 1 : 500 (2.19 ug/ml) dilution; PD1 with ab237728 at 1 : 2000 (0.525 ug/ml) dilution and CD68 with ab213363 at 1 : 500 (1.26 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD68 (magenta; Opal^™690), anti-PD-L1 (green; Opal^™520) and anti-PD1 (red; Opal^™570) on human tonsil.
Panel B : anti-PD-L1 stained on cells involved in T cell inhibition.
Panel C : anti-PD1 stained on antigen-stimulated T cells.
Panel D : anti-CD68 stained on macrophages.

The section was incubated in three rounds of staining : in the order of ab213363 and ab205921 for 30 mins, then ab237728 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins