Anti-CD68 抗体 [KP1]

• IHC-P

AbReview20018****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [KP1] (AB955)

ab955 staining CD68 in human ulcerated oral (Mucosa/Bone) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton-X 100 in PBS and blocked with 2.5% serum for 90 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/500 in 1% serum in PBS +0.01% Triton-X 100) for 16 hours at 4°C. A commercial IHC kit and DAB was used to visualize the staining.

This image is courtesy of an anonymous Abreview

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [KP1] (AB955)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue labelling CD68 with ab955 at 1/3000 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab93684 a Goat Anti-mouse IgG H&L (HRP) was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasmic staining on macrophages of human tonsil (PMID : 19543531).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [KP1] (AB955)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue labelling CD68 with ab955 at 1/3000 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab93684 a Goat Anti-mouse IgG H&L (HRP) was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Cytoplasmic staining on Kupffer cells of human liver (PMID : 12118106).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [KP1] (AB955)

Immunocytochemistry analysis of THP-1 (human monocytic leukemia monocyte) cells labelling CD68 with ab955 at 1/50 dilution. Cells were fixed with 100% methanol. Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) (ab150113) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-Tubulin antibody (rabbit mAb) ab179504 - AlexaFluor^®594 Goat anti- Rabbit secondary ab150080 at 1/500 dilution (red). Nuclear DNA was labelled with DAPI (blue).

Confocal image showing cytoplasmic staining in THP-1 cells

• IHC-P

AbReview22099****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [KP1] (AB955)

ab955 staining CD68 in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Tissue was fixed with paraformaldehyde. Samples were then blocked with 10% serum for 3 hours at 22°C followed by incubation with the primary antibody at a 1/100 dilution for 16 hours at 4°C. An Alexa-Fluor^® 568 conjugated goat anti-mouse polyclonal was used as secondary antibody at a 1/400 dilution.

Image courtesy of an anonymous Abreview.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [KP1] (AB955)

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling CD68 with ab955 at a concentration of 0.1 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.

ab955 Anti-CD68 antibody [KP1] 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)

• WB

Supplier Data

Western blot - Anti-CD68 antibody [KP1] (AB955)

Blocking and diluting buffer and concentration : 5% NFDM /TBST Exposure time : Lane1 : 60 seconds; Lane2 : 5 seconds; Lane3 : 180 seconds; Lane4 : 7 seconds ab303565 works better than ab955 in western blot testing. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab955 in western blot.

Lanes 1 and 3:

Western blot - Anti-CD68 antibody [KP1] (ab955) at 1/1000 dilution

Lanes 2 and 4:

Western blot - Anti-CD68 antibody [RM1031] (<a href='/products/primary-antibodies/cd68-antibody-rm1031-ab303565'>ab303565</a>) at 1/1000 dilution

Lanes 1 - 2:

Human spleen tissue lysate at 20 µg

Lanes 3 - 4:

THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 37 kDa

Observed band size: 110 kDa

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