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GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751181

GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Human

  • Green monkey

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human GWL aa 1-350.

Species Reactivity

Human, Green monkey

Validated Applications

WB, IF-Cell, IP

Target Molecular Weight

Predicted band size: 97 kDa

Positive Control

HeLa cell lysate, LNCaP cell lysate, HepG2 cell lysate, MDA-MB-231 cell lysate, COS-1 cell lysate, HeLa.

Conjugation

unconjugated

Clone Number

PSH07-69

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:50

  • IP

  • 1-2μg/sample

Target

Function

Serine/threonine kinase that plays a key role in M phase by acting as a regulator of mitosis entry and maintenance. Acts by promoting the inactivation of protein phosphatase 2A (PP2A) during M phase: does not directly inhibit PP2A but acts by mediating phosphorylation and subsequent activation of ARPP19 and ENSA at 'Ser-62' and 'Ser-67', respectively. ARPP19 and ENSA are phosphatase inhibitors that specifically inhibit the PPP2R2D (PR55-delta) subunit of PP2A. Inactivation of PP2A during M phase is essential to keep cyclin-B1-CDK1 activity high. Following DNA damage, it is also involved in checkpoint recovery by being inhibited. Phosphorylates histone protein in vitro; however such activity is unsure in vivo. May be involved in megakaryocyte differentiation.

Background References

1. Gall-Duncan T et al. Antagonistic roles of canonical and Alternative-RPA in disease-associated tandem CAG repeat instability. Cell. 2023 Oct.

2. Yang J et al. Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins. Cell Rep. 2023 Feb.

Subcellular Location

Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, Nucleus, Cleavage furrow.

UNIPROT

SwissProt: Q96GX5 Human

Synonyms

2700091H24Rik

C88295

FLJ14813

GREATWALL

Greatwall protein kinase

GW

GWL

GWL_HUMAN

hGWL

MAST-L

Expand

Images

  • Western blot analysis of GWL on different lysates with Rabbit anti-GWL antibody (<a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: LNCaP cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: MDA-MB-231 cell lysate<br />Lane 5: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 97 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 59 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/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GWL on different lysates with Rabbit anti-GWL antibody (HA751181) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: LNCaP cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: MDA-MB-231 cell lysate
    Lane 5: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 97 kDa
    Observed band size: 110 kDa

    Exposure time: 59 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 (HA751181) at 1/2,000 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.

  • Immunocytochemistry analysis of HeLa cells labeling GWL with Rabbit anti-GWL antibody (<a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GWL antibody (<a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a>) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. 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. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/50 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling GWL with Rabbit anti-GWL antibody (HA751181) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GWL antibody (HA751181) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/50 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • GWL was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs&reg; VHH(HRP) (<a href="/products/025-100-005" style="font-weight: bold;text-decoration: underline;">025-100-005</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751181" style="font-weight: bold;text-decoration: underline;">HA751181</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 40 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    GWL was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751181 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751181 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA751181 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA751181 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 40 seconds; ECL: K1802

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