Anti-MERTK antibody [EPR17534-139]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-MERTK antibody [EPR17534-139] (AB184086)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [EPR17534-139] (AB184086)

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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-MERTK antibody [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-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [EPR17534-139] (AB184086)

Immunohistochemical analysis of paraffin-embedded (A) Kidney tissue from wild-type C57BL/6JGpt mice / (B) Kidney tissue from MERTK knockout mice tissue labeling MERTK with ab184086 at 1/600 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Kidney tissue from wild-type C57BL/6JGpt mice and no staining on (B) Kidney tissue from MERTK knockout mice.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and MERTK-KO homozygous mice (Strain ID : T012765).

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-MERTK antibody [EPR17534-139] (AB184086)

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.

• IP

Supplier Data

Immunoprecipitation - Anti-MERTK antibody [EPR17534-139] (AB184086)

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).

All lanes:

Immunoprecipitation - Anti-MERTK antibody [EPR17534-139] (ab184086)

Predicted band size: 110 kDa

Observed band size: 110-140 kDa,180-210 kDa

false

• WB

Lab

Western blot - Anti-MERTK antibody [EPR17534-139] (AB184086)

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

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Mertk-KO homozygous mice (Strain ID : T012765).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was used at 1/200000 (36KDa).

All lanes:

Western blot - Anti-MERTK antibody [EPR17534-139] (ab184086) at 1/1000 dilution

Lane 1:

Mertk knockout mouse kidney (male) tissue lysate at 40 µg

Lane 2:

Wild-type mouse spleen tissue lysate (male) at 40 µg

Lane 3:

Mertk knockout mouse kidney (female) tissue lysate at 40 µg

Lane 4:

Wild-type mouse spleen tissue lysate (female) 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 1/100000 dilution

Observed band size: 110-150 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MERTK antibody [EPR17534-139] (AB184086)

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

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

All lanes:

Western blot - Anti-MERTK antibody [EPR17534-139] (ab184086) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 110 kDa

Observed band size: 110-140 kDa,180-210 kDa

false

Exposure time: 3min