Anti-MERTK 抗体 [EPR17534-139] (ab184086)

IHC image of MERTK staining in a section of frozen mouse normal liver performed on a Leica Biosystems BOND^® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab184086, 0.5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of MERTK staining in a section of frozen mouse normal spleen performed on a Leica Biosystems BOND^® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab184086, 0.1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

The expression profile observed is consistent with what has been described in the literature (PMID: 17047157).

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

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MERTK with ab184086 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous staining on mouse spleen (PMID: 19631584) is observed. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MERTK with ab184086 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on hepatic sinusoids of mouse liver (PMID: 23799121) is observed. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

MERTK was immunoprecipitated from 10 μg of mouse spleen tissue lysate with ab184086 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184086 at 1/1000 dilution. Secondary used is VeriBlot for IP Detection Reagent (HRP) (ab131366) at a 1/10,000 dilution.

Lane 1: Mouse spleen tissue lysate, 10 μg (input).
Lane 2: ab184086 IP in mouse spleen tissue lysate (+)
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184086 in mouse spleen lysate.

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

The molecular masses observed are consistent with what has been described in the literature. The band at approximately 50 kDa likely represents a cleavage fragment (PMID: 17047157, 15673687).