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MELK Recombinant Rabbit Monoclonal Antibody [PSH03-55]

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Specification

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Catalog# HA722014

MELK Recombinant Rabbit Monoclonal Antibody [PSH03-55]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

MELK Recombinant Rabbit Monoclonal Antibody [PSH03-55]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human MELK aa 1-400 / 651.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 75 kDa

Positive Control

HeLa cell lysate, 293T cell lysate, MDA-MB-231 cell lysate, HCT 116 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, human breast cancer tissue, human testis tissue.

Conjugation

unconjugated

Clone Number

PSH03-55

RRID

AB_3096635

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:500-1:2,000

Target

Function

Maternal embryonic leucine zipper kinase (MELK) is an enzyme that in humans is encoded by the MELK gene. MELK is a serine/threonine kinase belonging to the family of AMPK/Snf1 protein kinases. MELK was first identified present as maternal mRNA in mouse embryos. MELK expression is elevated in a number of cancers and is an active research target for pharmacological inhibition. MELK was previously believed to be essential for cancer cell proliferation. However, recent research using CRISPR has demonstrated that MELK is fully dispensable for cancer cell growth, casting doubt on the rationale for targeting this protein in patients. The results are dependent on the experimental design. Therefore, there is a need for further research. MELK has been shown to interact with CDC25B.

Background References

1. Tang B et al. Pharmacological inhibition of MELK restricts ferroptosis and the inflammatory response in colitis and colitis-propelled carcinogenesis. Free Radic Biol Med. 2021 Aug

2. McDonald IM et al. Enigmatic MELK: The controversy surrounding its complex role in cancer. J Biol Chem. 2020 Jun

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q14680 Human

SwissProt: Q61846 Mouse

Synonyms

AI327312 antibody

hMELK antibody

HPK 38 antibody

hPK38 antibody

KIAA0175 antibody

Likely ortholog of maternal embryonic leucine zipper kinase antibody

Maternal embryonic leucine zipper kinase antibody

MELK antibody

MELK_HUMAN antibody

mKIAA0175 antibody

Expand

Images

  • Western blot analysis of MELK on different lysates with Rabbit anti-MELK antibody (<a href="/products/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: 293T cell lysate<br />Lane 3: MDA-MB-231 cell lysate<br />Lane 4: HCT 116 cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: C2C12 cell lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 75 kDa<br />Observed band size: 72 kDa<br /><br />Exposure time: 2 minutes;<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/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 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 1/100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MELK on different lysates with Rabbit anti-MELK antibody (HA722014) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: 293T cell lysate
    Lane 3: MDA-MB-231 cell lysate
    Lane 4: HCT 116 cell lysate
    Lane 5: NIH/3T3 cell lysate
    Lane 6: C2C12 cell lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 75 kDa
    Observed band size: 72 kDa

    Exposure time: 2 minutes;

    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 (HA722014) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MELK on different lysates with Rabbit anti-MELK antibody (<a href="/products/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 dilution.<br /><br />Lane 1: Hepa1-6-si NT cell lysate<br />Lane 2: Hepa1-6-si MELK cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 75 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 20 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/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 MELK on different lysates with Rabbit anti-MELK antibody (HA722014) at 1/2,000 dilution.

    Lane 1: Hepa1-6-si NT cell lysate
    Lane 2: Hepa1-6-si MELK cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 75 kDa
    Observed band size: 75 kDa

    Exposure time: 20 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 (HA722014) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 breast cancer tissue with Rabbit anti-MELK antibody (<a href="/products/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/500 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/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/500 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 breast cancer tissue with Rabbit anti-MELK antibody (HA722014) at 1/500 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 (HA722014) at 1/500 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 testis tissue with Rabbit anti-MELK antibody (<a href="/products/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 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/HA722014" style="font-weight: bold;text-decoration: underline;">HA722014</a>) at 1/2,000 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 testis tissue with Rabbit anti-MELK antibody (HA722014) at 1/2,000 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 (HA722014) at 1/2,000 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation