Anti-CD18 antibody [EPR26929-26]

• Flow Cyt

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Flow Cytometry - Anti-CD18 antibody [EPR26929-26] (AB307406)

Flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell, Left) / U937 (human histiocytic lymphoma monocyte, Right) cells labelling CD18 with ab307406 at 1/50 dilution (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). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Negative control : HeLa (PMID : 31109948).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD18 antibody [EPR26929-26] (AB307406)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD18 with ab307406 at 1/100 (5.22 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on human spleen. The section was incubated with ab307406 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD18 antibody [EPR26929-26] (AB307406)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CD18 with ab307406 at 1/100 (5.22 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Negative control : no staining on human cardiac muscle. The section was incubated with ab307406 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD18 antibody [EPR26929-26] (AB307406)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling CD18 with ab307406 at 1/100 (5.22 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on (A) HEK-293T transfected with a ITGB2 his tag construct, no staining on (B) HEK-293T transfected with empty plasmid. The section was incubated with ab307406 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD18 antibody [EPR26929-26] (AB307406)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-937 (human histiocytic lymphoma monocyte) cells labelling CD18 with ab307406 at 1/1000 (0.522 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 U-937 cell line. Negative control : HeLa. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8) 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD18 antibody [EPR26929-26] (AB307406)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling CD18 with ab307406 at 1/100 (5.22 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on macrophages of human lung. The section was incubated with ab307406 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-CD18 antibody [EPR26929-26] (AB307406)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD18 with ab307406 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Cells were stained with Rabbit IgG or ab307406. Then stained with anti-CD4 conjugated to Alexa Fluor^® 647.

• Flow Cyt

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Flow Cytometry - Anti-CD18 antibody [EPR26929-26] (AB307406)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells lABelling CD18 with ab307406 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Cells were stained with Rabbit IgG or ab307406. Then stained with anti-CD14 conjugated to Alexa Fluor^® 647.

• WB

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Western blot - Anti-CD18 antibody [EPR26929-26] (AB307406)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : Hela The MW is consistent to what has been described in the literature (PMID : 31109948) Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 26 seconds

All lanes:

Western blot - Anti-CD18 antibody [EPR26929-26] (ab307406) at 1/1000 dilution

Lane 1:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate 20 μg

Lane 2:

Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 μg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 90 kDa,110 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-CD18 antibody [EPR26929-26] (AB307406)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : human heart The MW is consistent to what has been described in the literature (PMID : 31109948) Samples are non-boiled as boiling may cause protein aggregation. Exposure time : Lanes 1 : 37 seconds, Lanes 2-3 : 59 seconds, Lanes 4-5 : 103 seconds,

All lanes:

Western blot - Anti-CD18 antibody [EPR26929-26] (ab307406) at 1/1000 dilution

Lane 1:

Human lymphoma tissue 20 μg

Lane 2:

Human heart tissue 20 μg

Secondary

All lanes:

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

Observed band size: 90 kDa,110 kDa

false

Exposure time: 37s