Anti-YKL-40/CHI3L1 antibody [EPR27396-56]

• IHC

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Immunohistochemistry - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling YKL-40/CHI3L1 with ab305035 at 1/500 (1.092 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human glioma. The section was incubated with ab305035 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

• ICC

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Immunocytochemistry - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling YKL-40/CHI3L1 with ab305035 at 1/50 (10.92 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in THP-1 cell line.Negative control : A-204.The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A-204 (human muscle rhabdomyosarcoma, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling YKL-40/CHI3L1 with ab305035 at 1/500 dilution (0.1ug) (Red) (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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : A-204.

• IHC

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Immunohistochemistry - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling YKL-40/CHI3L1 with ab305035 at 1/500 (1.092 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human spleen. The section was incubated with ab305035 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-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (ab305035) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 20 µg

Lane 2:

Human spleen 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

Predicted band size: 43 kDa

Observed band size: 42 kDa

true

Exposure time: 180s

• WB

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Western blot - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human CHIT1 or CHI3L2. Exposure time : 8 seconds

All lanes:

Western blot - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (ab305035) at 1/1000 dilution

Lane 1:

Human CHI3L1 recombinant protein (No tag, aa22-383) at 10 ng

Lane 2:

293T transfected with human CHITI (aa 22-466) expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T transfected with human CHI3L2 (aa 27-390) 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

Predicted band size: 43 kDa

Observed band size: 42 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (AB305035)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative controls : A-204, Jurkat (PMID : 16472595) Exposure time : 48 seconds

All lanes:

Western blot - Anti-YKL-40/CHI3L1 antibody [EPR27396-56] (ab305035) at 1/1000 dilution

Lane 1:

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

Lane 2:

A-204 (human muscle rhabdomyosarcoma cell), whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), 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

Predicted band size: 43 kDa

Observed band size: 42 kDa

false

Exposure time: 48s