Anti-Langerin antibody [EPR28089-89]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR28089-89] (AB317813)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Langerin with ab317813 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver.
The section was incubated with ab317813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Langerin antibody [EPR28089-89] (AB317813)

Multiplex immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections of mouse skin tissue staining with ab309143 at 1/100 dilution, ab317813 at 1/500 dilution, and ab315806 at 1/500 dilution. The secondary used was Opal Polymer HRP Ms + Rb. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Panel A : merged staining of anti-Collagen VII (green; Opal™520), anti-Langerin (gray; Opal™570) and anti-Loricrin (magenta; Opal™690) on mouse skin. Panel B : anti-Collagen VII staining the dermal–epidermal junction in mouse skin. Panel C : anti-Langerin staining Langerhans cells in mouse skin. Panel D : anti-Loricrin staining terminally differentiated epidermal 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 ab309143, ab317813 and ab315806 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR28089-89] (AB317813)

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling Langerin with ab317813 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Langerhans cells of mouse skin.
The section was incubated with ab317813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Langerin antibody [EPR28089-89] (AB317813)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Langerin with ab317813 at 1/500 (1.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab317813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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

• IP

Supplier Data

Immunoprecipitation - Anti-Langerin antibody [EPR28089-89] (AB317813)

Langerin was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab317813 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317813 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse spleen tissue lysate
Lane 2 : ab317813 IP in Mouse spleen tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317813 in mouse spleen tissue lysate

All lanes:

Immunoprecipitation - Anti-Langerin antibody [EPR28089-89] (ab317813) at 1/30 dilution

All lanes:

Mouse spleen tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Langerin antibody [EPR28089-89] (AB317813)

Negative control : Mouse liver and kidney

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

All lanes:

Western blot - Anti-Langerin antibody [EPR28089-89] (ab317813) at 1/1000 dilution

Lane 1:

Mouse lymph node tissue lysate at 20 µg

Lane 2:

Mouse thymus tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Mouse kidney 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 1/100000 dilution

Observed band size: 40-50 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Langerin antibody [EPR28089-89] (AB317813)

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

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

All lanes:

Western blot - Anti-Langerin antibody [EPR28089-89] (ab317813) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse CD207 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: 40-50 kDa,36 kDa

false

Exposure time: 15s