Anti-MERTK antibody [EPR26359-12]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-MERTK antibody [EPR26359-12] (AB300136)

Flow cytometric analysis of HL-60 (human acute promyelocytic leukemia promyeloblast, Left) / HepG2 (human hepatocellular carcinoma epithelial cell, Right) cells labelling MERTK with ab300136 at 1/50 dilution (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® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Negative control : HL-60 (PMID : 23474756).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-MERTK antibody [EPR26359-12] (AB300136)

Flow cytometric analysis of WEHI-231 (mouse B cell lymphoma B lymphocyte, Left) / J774A.1 (mouse reticulum cell sarcoma monocyte macrophage, Right) cells labelling MERTK with ab300136 at 1/50 dilution (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). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Negative control : WEHI-231(PMID : 16652142; PMID : 12884290)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MERTK antibody [EPR26359-12] (AB300136)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized J774A.1(mouse reticulum cell sarcoma monocyte macrophage) cells labeling MERTK with ab300136 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in J774A.1 cell line. Negative control : WEHI-231(PMID : 16652142; PMID : 12884290).

Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 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) preabsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MERTK antibody [EPR26359-12] (AB300136)

Immunohistochemical analysis of paraffin-embedded (A) Kidney tissue from wild-type C57BL/6J mice and (B) Kidney tissue from MERTK knockout mice tissue labeling MERTK with ab300136 at 1/1000 dilution.

Positive staining on (A) Kidney tissue from wild-type C57BL/6J mice, 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. Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

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

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-MERTK antibody [EPR26359-12] (AB300136)

Flow cytometric analysis of NR8383 (rat lung macrophage alveolar) cells labelling MERTK with ab300136 at 1/50 dilution (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). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• WB

Lab

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : ECL western blotting substrate 180 seconds, higher sensitivity ECL western substrate 80 seconds.

The left image uses the ECL western blotting substrate and the right image uses the higher sensitivity ECL western substrate.

We recommend using a higher sensitive ECL substrate to increase the band intensity.

All lanes:

Western blot - Anti-MERTK antibody [EPR26359-12] (ab300136) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), 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 1/20000 dilution

Predicted band size: 110 kDa

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

false

• WB

Supplier Data

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

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

Negative control : HL-60 (PMID : 23474756)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16652142).

All lanes:

Western blot - Anti-MERTK antibody [EPR26359-12] (ab300136) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HL-60 (human acute promyelocytic leukemia promyeloblast) 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

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

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

ab52968 was used at 1/1000 dilution. ab184086 was used at 1/1000 dilution. ab181602 was used as a loading control at 1/1000000 dilution. Blocking/Diluting buffer and concentration : 5% NFDM/TBST For Western Blot, it is recommended to use ab52968 for detecting human samples and ab184086 for detecting mouse samples.

All lanes:

Western blot - Anti-MERTK antibody [EPR26359-12] (ab300136) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) 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 1/20000 dilution

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

false

• WB

Lab

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

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

All lanes:

Western blot - Anti-MERTK antibody [EPR26359-12] (ab300136) at 1/1000 dilution

Lane 1:

Wild-type mouse kidney tissue lysate at 40 µg

Lane 2:

Mertk knockout mouse kidney (case 1) tissue lysate at 40 µg

Lane 3:

Mertk knockout mouse kidney (case 2) tissue 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: 110-140 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling MERTK with ab300136 at 1/800 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Membranous staining on mouse spleen (PMID : 25479139) is observed. The section was incubated with ab300136 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• WB

Supplier Data

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labelling MERTK with ab300136 at 1/800 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Membranous staining on endothelial cells of mouse kidney (PMID : 29350876) is observed. The section was incubated with ab300136 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Supplier Data

Western blot - Anti-MERTK antibody [EPR26359-12] (AB300136)

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

Negative control : WEHI-231 (PMID : 12884290).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16652142).

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-MERTK antibody [EPR26359-12] (ab300136) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 2:

WEHI-231 (mouse B cell lymphoma B 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

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

false

Exposure time: 92s