Anti-CD68 抗体 [EPR23917-164]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling CD68 with ab283654 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Flow cytometry staining of C57BL/6 mouse splenocytes (top) or C57BL/6 Mouse Bone Marrow Cell-derived Macrophages (bottom) with ab283654 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Cells were incubated for 30min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab283654 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶in 100 μl at 1.0 μg/ml (1/2212)) for 30min at 22°C. The cells were simultaneously stained with F4/80.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh) tissue labeling CD68 with ab283654 at 1/100 (4.66 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on macrophage of rat spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse small intestine tissue staining CD68 with ab283654 at a 1/100 dilution, ab245235 anti-CD19 used at 1/1000 dilution and ab237721 anti-CD3 used at a 1/2000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD68 (magenta; Opal^™520), anti-CD19 (green; Opal^™690) and anti-CD3 (grey; Opal^™570) on mouse small intestine.
Panel B : anti-CD68 staining macrophages in mouse small intestine.
Panel C : anti-CD19 staining B lymphocytes in mouse small intestine.
Panel D : anti-CD3 staining T lymphocytes in mouse small intestine.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab283654, ab245235 and ab237721 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining CD68 with ab283654 at a 1/100 dilution, ab245235 anti-CD19 used at 1/1000 dilution and ab237721 anti-CD3 used at a 1/2000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD68 (magenta; Opal^™520), anti-CD19 (green; Opal^™690) and anti-CD3 (grey; Opal^™570) on mouse colon.
Panel B : anti-CD68 staining macrophages in mouse colon.
Panel C : anti-CD19 staining B lymphocytes in mouse colon.
Panel D : anti-CD3 staining T lymphocytes in mouse colon.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab283654, ab245235 and ab237721 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung treated with lps (1ug/ml 16h)+bfa (1ug/ml 16h) (+) ex vivo tissue staining Pentraxin 3/PTX3 with ab325639 at a 1 : 100 (5.07 µg/ml) dilution, ab283654 anti-CD68 used at 1 : 100 (4.15 µg/ml) dilution.

Panel A : merged staining of anti-Pentraxin 3 (green; Opal™520), anti-E CD68 (magenta; Opal™570) on mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel B : anti-Pentraxin 3 staining stroma cells in mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel C : anti-CD68 staining macrophages in mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325639 and ab283654 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.

Nuclear counter stain with DAPI.

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 [EPR23917-164] (AB283654)

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Paraffin embedded mouse spleen tissue slides were dewaxed, followed by heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, Epitope Retrieval ER2 Solution) for 20 mins. Endogenous peroxidase activity was blocked by Refine Detection Kit Peroxide Block for 10 mins. The sections were then incubated with ab283654 (1/100) at room temperature for 30 mins, followed by a ready to use LeicaDS9800 (BOND Polymer Refine Detection Kit). The sections were counterstained with Hematoxylin. The images were taken with a Leica Aperio AT2. ab283654 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in IHC-P. The image shows the staining intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung tissue staining CD62L with ab325322 at a 1/1000 (0.509 μg/ml) dilution, ab283654 anti-CD68 used at a 1/100 (4.43 μg/ml) dilution and ab324911 anti-ATP-binding cassette sub-family A member 3 used at a 1/2000 (0.253 μg/ml) dilution.

Panel A : anti-CD62L (green; Opal™520), anti-CD68 (magenta; Opal™690), anti-ATP-binding cassette sub-family A member 3 (gray; Opal™570) on mouse lung.

Panel B : anti-CD62L staining immune cells in mouse lung.

Panel C : anti-CD68 staining macrophages in mouse lung.

Panel D : anti-ATP-binding cassette sub-family A member 3 staining type Ⅱ pneumocytes in mouse lung.

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

The section was incubated in three rounds of staining : in the order of ab325322, ab283654 and ab324911 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat liver tissue staining MARCO with ab323717 at a 1 : 200 (2.49 ug/ml) dilution, ab283654 anti-CD68 used at 1 : 100 (4.43 ug/ml) dilution and ab237721 anti-CD3 used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MARCO (green; Opal™520), anti-CD68 (magenta; Opal™690) and anti-CD3 (yellow; Opal™570) on rat liver.
Panel B : anti-MARCO staining macrophages in rat liver.
Panel C : ant-CD68 staining macrophages in rat liver.
Panel D : ant-CD3 staining T lymphocytes in rat liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323717, ab283654 and ab237721 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.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung tissue staining MARCO with ab323717 at a 1 : 200 (2.49 ug/ml) dilution, ab283654 anti-CD68 used at 1 : 100 (4.43 ug/ml) dilution and ab237721 anti-CD3 used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MARCO (green; Opal™520), anti-CD68 (magenta; Opal™690) and anti-CD3 (yellow; Opal™570) on mouse lung.
Panel B : anti-MARCO staining macrophages in mouse lung.
Panel C : ant-CD68 staining macrophages in mouse lung.
Panel D : ant-CD3 staining T lymphocytes in mouse lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323717, ab283654 and ab237721 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.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining CD5L/CT-2 with ab324193 at a 1 : 500 (0.958 ug/ml) dilution, ab283654 anti-CD68 used at 1 : 100 (4.43 ug/ml) dilution and ab237721 anti-CD3 used at a 1 : 2000 (0.25 ug/ml) dilution.

Panel A : merged staining of anti-CD5 antigen-like (green; Opal™520), anti-CD68 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse liver.
Panel B : anti-CD5 antigen-like staining macrophages in mouse liver.
Panel C : anti-CD68 staining macrophages in mouse liver.
Panel D : anti-CD3 staining T lymphocytes in mouse liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324193, ab283654 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Nuclear counter stain with DAPI.

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

• IHC

Lab

Immunohistochemistry - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of formalin fixed paraffin embedded mouse spleen labelling CD68 with ab283654 at a concentration of 0.1µ/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.

ab283654 anti-CD68 antibody [EPR23917-164] 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)

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling CD68 with ab283654 at 1/100 (4.66 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on Kupffer cells of rat liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells in mouse colon. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 cells labelling CD68 with ab283654 at 1/50 (9.32 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in J774A.1 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on macrophages in rat lung. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat spleen. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Kupffer cells in rat liver. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (AB283654)

Immunohistochemical analysis of paraffin-embedded Mouse large B cell lymphoma tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse large B cell lymphoma. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

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

• WB

Lab

Western blot - Anti-CD68 antibody [EPR23917-164] (AB283654)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of mouse spleen lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab283654 (1/5000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/50,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab283654 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

All lanes:

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/5000 dilution

All lanes:

Mouse spleen lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD68 antibody [EPR23917-164] (AB283654)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1-2 : Merged signal (red and green). Green - ab283654 observed at 100 kDa. Red - loading control ab8245 observed at 36 kDa.
ab283654 Anti-CD68 antibody [EPR23917-164] was shown to specifically react with CD68 in wild-type RAW264.7 cells. Loss of signal was observed when knockout cell line (knockout cell lysate - ab280106) was used. Wild-type and CD68 knockout samples were subjected to SDS-PAGE. ab283654 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution

Lane 1:

Wild-type RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

Western blot - Mouse CD68 knockout RAW 264.7 cell lysate (<a href='/products/cell-lysates/mouse-cd68-knockout-raw-2647-cell-lysate-ab280106'>ab280106</a>) at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 37 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-CD68 antibody [EPR23917-164] (AB283654)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : mouse skeletal muscle (PMID : 28091823).

Exposure time : 37 seconds

All lanes:

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 37 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-CD68 antibody [EPR23917-164] (AB283654)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : Lane 1 : 7.75 seconds; Lane 2 : 125 seconds; Lane 3 : 92 seconds

All lanes:

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate 20 at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 37 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-CD68 antibody [EPR23917-164] (AB283654)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Macrosialin (CD68) is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID : 7680921)

Exposure time : Lane 1-2 : 7.75 seconds; Lane 3-4 : 48 seconds

All lanes:

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 15 µg

Lane 2:

RAW264.7 treated with PNGase F whole cell lysate at 15 µg

Lane 3:

Mouse spleen tissue lysate at 15 µg

Lane 4:

Mouse spleen treated with PNGase F tissue lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 37 kDa

Observed band size: 100 kDa

false