Anti-Ly6c antibody [EPR27220-23]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Immunohistochemical analysis of paraffin-embedded Panel A : HEK-293T ( tissue labeling Ly6c with ab314120 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) HEK-293T transfected with mouseLY6C1 expression vector and (B) HEK-293T cells transfected with a mouse LY6C2 protein expression vector, no staining on (C) HEK-293T cells transfected with a mouse Ly6i protein expression vector, (D) HEK-293T cells transfected with a mouse Ly6g protein expression vector, (E) HEK-293T cells transfected with a mouse Ly6a protein expression vector and (F) HEK-293T transfected with empty vector.The section was incubated with ab314120 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Ly6c with ab314120 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse colon vascular endothelium. The section was incubated with ab314120 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Ly6c with ab314120 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse spleen. The section was incubated with ab314120 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.

• mIHC

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Multiplex immunohistochemistry - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue staining Ly6c with ab314120 at a 1/100 dilution ( 5.19 μg/ml), ab245235 anti-CD19 used at 1/1000 dilution (0.999 μg/ml), ab133357 anti-CD11b used at 1/20000 dilution (0.05 μg/ml). Panel A : merged staining of anti-Ly6c (green; Opal™690), anti-CD19 (gray; Opal™520) and anti-CD11b (red; Opal™570) on mouse spleen. Panel B : anti-Ly6c stained on monocytes/macrophages. Panel C : anti-CD19 stained on B cells. Panel D : anti-CD11b stained on monocytes/macrophages. The section was incubated in three rounds of staining : in the order of ab314120, ab245235, and ab133357 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins 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-Ly6c antibody [EPR27220-23] (AB314120)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Ly6c with ab314120 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse cerebrum vascular endothelium. The section was incubated with ab314120 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

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Western blot - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 92 seconds

All lanes:

Western blot - Anti-Ly6c antibody [EPR27220-23] (ab314120) at 1/1000 dilution

Lane 1:

Mouse bone marrow lysate at 20 µg

Lane 2:

Mouse kidney lysate at 20 µg

Lane 3:

Mouse heart 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: 14 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : testis, liver In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 92 seconds

All lanes:

Western blot - Anti-Ly6c antibody [EPR27220-23] (ab314120) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 20 µg

Lane 2:

Mouse spleen lysate at 20 µg

Lane 3:

Mouse testis lysate at 20 µg

Lane 4:

Mouse liver 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: 14 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-Ly6c antibody [EPR27220-23] (AB314120)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody recognizes both Ly6c1 and Ly6c2. 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 Exposure time : 15 seconds

All lanes:

Western blot - Anti-Ly6c antibody [EPR27220-23] (ab314120) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with a mouse Ly6c1 expression vector containing a HA-tag whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a mouse Ly6c2 expression vector containing a His-tag whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a mouse Ly6i expression vector containing a His-tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse Ly6g expression vector containing a His-tag whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a mouse Ly6a 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: 14 kDa

false

Exposure time: 15s