Anti-Langerin antibody [EPR24685-12]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Langerin with ab283686 at 1/500 (1.066 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on human cerebrum. The section was incubated with ab283686 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized transfected HEK-293T cells labelling Langerin with ab283686 at 1/1000 (1.008 μg/ml) followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing positive staining in HEK-293T cells transfected with a human CD207/Langerin expression vector containing a myc-His-tag® (shown in green). ab202008 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 488) at 1/100 (5μg/ml) was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labelling Langerin with ab283686 at 1/500 (1.066 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on scattered cells in human lung carcinoma. The section was incubated with ab283686 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 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-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labelling Langerin with ab283686 at 1/500 (1.066 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on Langerhans cells in human skin. The section was incubated with ab283686 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Flow cytometry (intracellular) analysis of transfected 293T cells. Cells were fixed with 4% PFA for 10 minutes and permeabilised with 0.1% Tween-20 before staining for Langerin in 293T cells transfected with a human Langerin expression vector containing a myc-His-tag® using ab283686 1/1000 dilution (0.1ug) conjugated to Alexa Fluor® 647. (Middle panel). Staining was compared with 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right panel) and with an isotype control, Rabbit monoclonal IgG (ab172730) (left panel). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody at a 1/5000 dilution.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Langerin antibody [EPR24685-12] (AB283686)

Panel A : merged staining of anti-Desmoglein 1/DSG1 (gray; Opal™690), anti-Langerin (green; Opal™520) and anti-TRP2/DCT (red; Opal™570) on human skin. Panel B : anti-TRP2/DCT stained on melanocytes. Panel C : anti-Langerin stained on Langerhans cells. Panel D : anti-Desmoglein 1/DSG1 stained on epidermis. The section was incubated in three rounds of staining : in the order of ab236259, ab283686, and ab221144 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-Langerin antibody [EPR24685-12] (AB283686)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse skin staining of Cytokeratin 1 + Cytokeratin 2e with ab315228 at a 1/2000 (0.263 μg/ml) dilution, Collagen I with ab270993 and Langerin with ab283686 at a 1/1000 dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Cytokeratin 1 + Cytokeratin 2e (green; Opal™520), anti-Collagen I (magenta; Opal™690) and anti-Langerin (gray; Opal™570) on mouse skin.
Panel B : anti-Cytokeratin 1 + Cytokeratin 2e staining epidermis keratin layer in mouse skin.
Panel C : anti-Collagen I staining connective tissues in mouse skin.
Panel D : anti-Langerin staining Langerhans cells in mouse skin.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315228, ab270993 and ab283686 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system,

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized transfected HEK-293T cells labelling Langerin with ab283686 at 1/1000 (1.008 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing positive staining in HEK-293T cells transfected with a mouse CD207/Langerin expression vector containing a myc-His-tag® (shown in green). ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) at 1/100 (5μg/ml) was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse thymus (fresh) tissue labeling Langerin with ab283686 at 1/100 (5.33 μg/mL) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green). Positive staining on mouse thymus is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 μg/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labelling Langerin with ab283686 at 1/500 (1.066 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on Langerhans cells in mouse skin. The section was incubated with ab283686 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 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-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of paraffin-embedded Mouse thymus tissue labelling Langerin with ab283686 at 1/500 (1.066 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on scattered immune cells in mouse thymic medulla. The section was incubated with ab283686 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Langerin antibody [EPR24685-12] (AB283686)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skin (fresh) tissue labeling Langerin with ab283686 at 1/100 (5.33 μg/mL) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green). Positive staining on Langerhans cells of mouse skin is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 μg/mL dilution.

• WB

Lab

Western blot - Anti-Langerin antibody [EPR24685-12] (AB283686)

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

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

In Western blot, Anti-6X His tag® antibody [AD1.1.10] total protein control (1/1000) (30KDa)

All lanes:

Western blot - Anti-Langerin antibody [EPR24685-12] (ab283686) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human CD207/Langerin expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with a mouse CD207/Langerin expression vector containing a His-tag, whole cell lysate (Low Expression) 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: 40 kDa

false

Exposure time: 15s