Anti-CD1d antibody [EPR29917-692]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD1d antibody [EPR29917-692] (AB324952)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling CD1d with ab324952 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in Jurkat cells (shown in green) . The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : K-562.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324952 at 1/50, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD1d antibody [EPR29917-692] (AB324952)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD1d with ab324952 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).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with anti-CD19 conjugated to PE/CY7.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD1d antibody [EPR29917-692] (AB324952)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD1d with ab324952 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).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with anti-CD3 conjugated to Brilliant Violet 421.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD1d antibody [EPR29917-692] (AB324952)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling CD1d with ab324952 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in subsets of human CD3 negative PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-rat CD3 mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 dilution (Magenta). 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 dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD1d antibody [EPR29917-692] (AB324952)

Flow cytometric analysis of K-562 (human chronic myelogenous leukemia lymphoblast, Left) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD1d with ab324952 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).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Negative control : K-562 (PMID : 32324315).

• WB

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Western blot - Anti-CD1d antibody [EPR29917-692] (AB324952)

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

This antibody does not cross-react with overexpressed human CD1C, CD1A or CD1B.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-CD1d antibody [EPR29917-692] (ab324952) at 1/1000 dilution

Lane 1:

LIF/His-tagged human CD1D recombinant protein at 10 ng

Lane 2:

LIF/His-tagged human CD1C recombinant protein at 10 ng

Lane 3:

293T cells transfected with a mouse CD1A expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse CD1B expression vector containing a His-tag, whole cell lysate at 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,55 kDa

false

Exposure time: 15s