Anti-YKL-40/CHI3L1 antibody [RM1204]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte) (Left) / CHI3L1 KO THP-1(FR090372) (Right) cells labelling YKL-40/CHI3L1 with ab319164 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) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of rat lung. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Mouse colon cancer tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse colon cancer. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse lung. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling YKL-40/CHI3L1 with ab319164 at 1/500 (1.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab319164 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

MSD279 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-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Negative control : 293T, Jurkat (PMID : 19767768, PMID : 16472595).

The molecular weight observed is consistent with what has been described in the literature (PMID : 19414556).

The identity of the lower MW band at approximately 25 kDa (in lane 3 and 5) is unknown.

In Western blot, ab319164 was shown to bind specifically to CHI3L1. Target of interest was observed at 43 kDa in wild-type THP-1 cell lysates (lane 1) with no signal observed at this size in CHI3L1 knockout cell line (lane 2, knockout cell line ab280038).

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

Exposure time : lane 1-2 : 25 seconds, lane 3-5 : 125 seconds

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-YKL-40/CHI3L1 antibody [RM1204] (ab319164) at 1/1000 dilution

Lane 1:

Wide-type THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

CHI3L1 knockout THP-1 whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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 1/20000 dilution

Observed band size: 43 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-YKL-40/CHI3L1 antibody [RM1204] (AB319164)

Low expression tissue : liver (PMID : 31250532).

The molecular weight observed is consistent with what has been described in the literature (PMID : 19414556).

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

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-YKL-40/CHI3L1 antibody [RM1204] (ab319164) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse liver 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: 43 kDa,36 kDa

false

Exposure time: 37s