Anti-CLEC5A antibody [EPR28104-85]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscl tissue labeling CLEC5A with ab313337 at 1/100 (5.34 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse skeletal muscle.The section was incubated with ab313337 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CLEC5A with ab313337 at 1/100 (5.34 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen (PMID : 19251634).The section was incubated with ab313337 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

Immunohistochemical analysis of paraffin-embedded Mouse bone marrow tissue labeling CLEC5A with ab313337 at 1/100 (5.34 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse bone marrow (PMID : 19251634).The section was incubated with ab313337 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

• IP

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Immunoprecipitation - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

CLEC5A was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab313337 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313337 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : ab313337 IP in RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313337 in RAW264.7 whole cell lysate

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

Exposure time : 93 seconds

All lanes:

Immunoprecipitation - Anti-CLEC5A antibody [EPR28104-85] (ab313337) at 1/1000 dilution

All lanes:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell 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

• WB

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Western blot - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : EL4 (PMID : 10449773). Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 37 seconds

All lanes:

Western blot - Anti-CLEC5A antibody [EPR28104-85] (ab313337) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate at 40 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 40 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: 37 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CLEC5A antibody [EPR28104-85] (AB313337)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : skeletal muscle. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 3 seconds

All lanes:

Western blot - Anti-CLEC5A antibody [EPR28104-85] (ab313337) at 1/1000 dilution

Lane 1:

Mouse bone marrow tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 37 kDa

false

Exposure time: 3s