Anti-C10 antibody [EPR23475-105]

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-C10 antibody [EPR23475-105] (AB275025)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse colon tissue labeling C10 with ab275025 at 1/500 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Positive staining on mouse colon (PMID : 7579352). The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C10 antibody [EPR23475-105] (AB275025)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling C10 with ab275025 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on mouse lung (PMID : 14734772). The section was incubated with ab275025 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-C10 antibody [EPR23475-105] (AB275025)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse small intestine tissue labeling C10 with ab275025 at 1/500 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Positive staining on mouse small intestine (PMID : 7579352). The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C10 antibody [EPR23475-105] (AB275025)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling C10 with ab275025 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on mouse spleen (PMID : 15770051). The section was incubated with ab275025 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-C10 antibody [EPR23475-105] (AB275025)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized P388D1 (Mouse lymphoma monocyte; macrophage) (treated with 1ug/ml lipopolysaccharide (LPS) for 4h then add 300ng/ml Brefeldin A for 16h) (Red) / Untreated control (Green) cells labelling C10 with ab275025 at 1/500 dilution (0.1ug) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Unknown

Immunoprecipitation - Anti-C10 antibody [EPR23475-105] (AB275025)

C10 was immunoprecipitated from 0.35 mg Mouse small intestine tissue lysate with ab275025 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275025 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : Mouse small intestine tissue lysate 10 ug

Lane 2 : ab275025 IP in Mouse small intestine tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275025 in mouse small intestine tissue lysate

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

Exposure time : 3 minutes.

The expression profile observed is consistent with what has been described in the literature (PMID : 19812544).

All lanes:

Immunoprecipitation - Anti-C10 antibody [EPR23475-105] (ab275025)

Predicted band size: 13 kDa

Observed band size: 13 kDa

false

• WB

Unknown

Western blot - Anti-C10 antibody [EPR23475-105] (AB275025)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 8 seconds.

This product does not recognize CCL9.

All lanes:

Western blot - Anti-C10 antibody [EPR23475-105] (ab275025) at 1/1000 dilution

Lane 1:

His-tagged mouse C10 recombinant protein, 20ng

Lane 2:

His-tagged mouse MIP-1 gamma (CCL9) recombinant protein (aa 22-122), 50ng

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 13 kDa

false