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MERTK Recombinant Rabbit Monoclonal Antibody [SR29-07] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750036

MERTK Recombinant Rabbit Monoclonal Antibody [SR29-07] - BSA and Azide free

Application

  • WB

  • IHC-P

  • FC

  • IF-Tissue

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

MERTK Recombinant Rabbit Monoclonal Antibody [SR29-07] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human MERTK aa 1-118 / 999.

Species Reactivity

Human

Validated Applications

WB, IHC-P, FC, IF-Tissue

Target Molecular Weight

Predicted band size: 110 kDa

Positive Control

293T cell lysate, HepG2 cell lysate, human tonsil tissue, 293T.

Conjugation

unconjugated

Clone Number

SR29-07

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:50-1:1,000

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:100

Target

Function

MerTK, also called c-Mer, is a member of the Mer/Axl/Tyro3 receptor kinase family. It is a 984 residue transmembrane protein made up of one tyrosine kinase domain, two Fibronectin type-III domains and two immunoglobulin-like C2-type domains. MerTK is the mammalian ortholog of the chicken retroviral oncogene product v-Eyk. This protein plays a critical role in macrophage activation, platelet aggregation, clot stability and the efficient removal of apoptotic cells. Specifically, MerTK acts as a signaling molecule, triggering outer segment ingestion in the retinal pigment epithelium (RPE) phagocytic process. Evidence suggests that MerTK signals via interaction with phosphatidylinositol-specific phospholipase C 2 (PI-PLC 2). When the gene encoding for MerTK is mutated, the RPE phagocytosis pathway is disrupted and autosomal recessive retinitis pigmentosa (RP) may result, leading to degeneration of retinal photoreceptor cells.

Background References

1. Pierce AM & Keating AK TAM receptor tyrosine kinases: Expression, disease and oncogenesis in the central nervous system. Brain Res 1542:206-20 (2014).

2. Ranganathan V et al. Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide RNAs. Nat Commun 5:4516 (2014).

Sequence Similarity

Belongs to the protein kinase superfamily. Tyr protein kinase family. AXL/UFO subfamily.

Tissue Specificity

Not expressed in normal B- and T-lymphocytes but is expressed in numerous neoplastic B- and T-cell lines. Highly expressed in testis, ovary, prostate, lung, and kidney, with lower expression in spleen, small intestine, colon, and liver.

Post-translational Modification

Autophosphorylated on Tyr-749, Tyr-753 and Tyr-754 in the activation loop allowing full activity. Autophosphorylated on Tyr-872 leading to recruitment of downstream partners of the signaling cascade such as PLCG2 (By similarity).

Subcellular Location

Membrane.

UNIPROT

SwissProt: Q12866 Human

Synonyms

c MER antibody

c mer proto oncogene tyrosine kinase antibody

c-mer antibody

cMER antibody

cmer protooncogene tyrosine kinase antibody

Eyk antibody

MER antibody

MER receptor tyrosine kinase antibody

MERK antibody

MERPEN antibody

Expand

Images

  • Western blot analysis of MERTK on different lysates with Rabbit anti-MERTK antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: HepG2 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 110 kDa<br />Observed band size: 150-210 kDa<br /><br />Exposure time: 40 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MERTK on different lysates with Rabbit anti-MERTK antibody (HA750036) at 1/1,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: HepG2 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 110 kDa
    Observed band size: 150-210 kDa

    Exposure time: 40 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750036) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MERTK on different lysates with Rabbit anti-MERTK antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-MERTK KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 110 kDa<br />Observed band size: 150-210 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of MERTK on different lysates with Rabbit anti-MERTK antibody (HA750036) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-MERTK KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 110 kDa
    Observed band size: 150-210 kDa

    Exposure time: 3 minutes; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750036) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MERTK antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MERTK antibody (HA750036) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750036) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling MERTK with Rabbit anti-MERTK antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>) at 1/100 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling MERTK with Rabbit anti-MERTK antibody (HA750036) at 1/100 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750036, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Flow cytometric analysis of 293T cells labeling MERTK.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA750036" style="font-weight: bold;text-decoration: underline;">HA750036</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of 293T cells labeling MERTK.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA750036, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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