Anti-CD1d antibody [EPR28086-741]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD1d antibody [EPR28086-741] (AB325682)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD1d with ab325682 at 1/1000 (0.506 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse cerebrum (PMID : 23891738).

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD1d antibody [EPR28086-741] (AB325682)

Immunohistochemical analysis of paraffin-embedded (A) WEHI-3 (mouse leukemia lymphoblast) cell pellet and (B) J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cell pellet labeling CD1d with ab325682 at 1/500 (1.012 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) WEHI-3 cell pellet, weak staining on (B) J774A.1 cell pellet.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-CD1d antibody [EPR28086-741] (AB325682)

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

Low expression : mouse brain (PMID : 2460336), J774A.1 (PMID : 2460336).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CD1d antibody [EPR28086-741] (ab325682) at 1/1000 dilution

Lane 1:

WEHI-3 (mouse leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Mouse thymus tissue lysate at 20 µg

Lane 4:

Mouse brain tissue 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 1/20000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 10s

• Dot

Lab

Dot Blot - Anti-CD1d antibody [EPR28086-741] (AB325682)

Dot blot analysis of CD1d using ab325682 at 1 : 1000 (0.506 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : 293T cells transfected with an empty vector containing a his tag whole cell lysate

Lane 2 : 293T cells transfected with a mouse CD1d1 expression vector containing a His-tag whole cell lysate

Lane 3 : 293T cells transfected with a mouse CD1d2 expression vector containing a His-tag whole cell lysate

Exposure time : 180 seconds.

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

This antibody cross-reacts with mouse CD1d2.

All lanes:

Dot Blot - Anti-CD1d antibody [EPR28086-741] (ab325682) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his tag whole cell lysate

Lane 2:

293T cells transfected with a mouse CD1d1 expression vector containing a His-tag whole cell lysate

Lane 3:

293T cells transfected with a mouse CD1d2 expression vector containing a His-tag whole cell lysate

Secondary

All lanes:

Dot 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

false

Exposure time: 180s