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SPAK Recombinant Rabbit Monoclonal Antibody [JE53-64]

Usd: 385

Specification

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Catalog# ET7110-37

SPAK Recombinant Rabbit Monoclonal Antibody [JE53-64]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

SPAK Recombinant Rabbit Monoclonal Antibody [JE53-64]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human SPAK aa 361-545 / 545.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 59 kDa

Positive Control

Jurkat cell lysate, Mouse brain tissue lysate, Mouse testis tissue lysate, Rat brain tissue lysate, Rat testis tissue lysate, human testis tissue, mouse testis tissue, rat testis tissue, human prostate carcinoma tissue, human pancreas tissue.

Conjugation

unconjugated

Clone Number

JE53-64

RRID

AB_3071002

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:200-1:1,000

Target

Function

STE20/SPS1-related proline-alanine-rich protein kinase is an enzyme that in humans is encoded by the STK39 gene. This gene encodes a serine/threonine kinase that is thought to function in the cellular stress response pathway. The kinase is activated in response to hypotonic stress, leading to phosphorylation of several cation-chloride-coupled cotransporters. The catalytically active kinase specifically activates the p38 MAP kinase pathway, and its interaction with p38 decreases upon cellular stress, suggesting that this kinase may serve as an intermediate in the response to cellular stress. Some studies suggest that this gene might be linked to high blood pressure.

Background References

1. Zhang J. et. al. Pharmacological targeting of SPAK kinase in disorders of impaired epithelial transport. Expert Opin Ther Targets. 2017 Aug;21(8):795-804.

2. Grimm PR. et. al. Constitutively Active SPAK Causes Hyperkalemia by Activating NCC and Remodeling Distal Tubules. J Am Soc Nephrol. 2017 Sep;28(9):2597-2606.

Sequence Similarity

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.

Tissue Specificity

Predominantly expressed in brain and pancreas followed by heart, lung, kidney, skeletal muscle, liver, placenta and testis.

Post-translational Modification

Phosphorylated at Ser-309 by PRKCQ. Autophosphorylation at Thr-231 positively regulates its activity (By similarity).

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q9UEW8 Human

SwissProt: Q9Z1W9 Mouse

SwissProt: O88506 Rat

Synonyms

DCHT antibody

DKFZp686K05124 antibody

OTTHUMP00000165175 antibody

PASK antibody

Proline alanine rich STE20 related kinase antibody

Serine threonine kinase 39 (STE20/SPS1 homolog yeast) antibody

Serine threonine kinase 39 antibody

Serine/threonine protein kinase 39 antibody

Serine/threonine-protein kinase 39 antibody

Small intestine SPAK like kinase antibody

Expand

Images

  • Western blot analysis of SPAK on different lysates with Rabbit anti-SPAK antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>) at 1/5,000 dilution.<br /><br />Lane 1: Jurkat cell lysate (15 µg/Lane)<br />Lane 2: Mouse brain tissue lysate (15 µg/Lane)<br />Lane 3: Mouse testis tissue lysate (15 µg/Lane)<br />Lane 4: Rat brain tissue lysate (15 µg/Lane)<br />Lane 5: Rat testis tissue lysate (15 µg/Lane)<br /><br />Predicted band size: 59 kDa<br />Observed band size: 59 kDa<br /><br />Exposure time: 13 seconds; ECL: K1801;<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/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 SPAK on different lysates with Rabbit anti-SPAK antibody (ET7110-37) at 1/5,000 dilution.

    Lane 1: Jurkat cell lysate (15 µg/Lane)
    Lane 2: Mouse brain tissue lysate (15 µg/Lane)
    Lane 3: Mouse testis tissue lysate (15 µg/Lane)
    Lane 4: Rat brain tissue lysate (15 µg/Lane)
    Lane 5: Rat testis tissue lysate (15 µg/Lane)

    Predicted band size: 59 kDa
    Observed band size: 59 kDa

    Exposure time: 13 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 (ET7110-37) at 1/5,000 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 testis tissue with Rabbit anti-SPAK antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</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/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</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 testis tissue with Rabbit anti-SPAK antibody (ET7110-37) 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 (ET7110-37) 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 mouse testis tissue with Rabbit anti-SPAK antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>) at 1/200 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/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>) at 1/200 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 mouse testis tissue with Rabbit anti-SPAK antibody (ET7110-37) at 1/200 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 (ET7110-37) at 1/200 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 rat testis tissue with Rabbit anti-SPAK antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</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/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</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 rat testis tissue with Rabbit anti-SPAK antibody (ET7110-37) 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 (ET7110-37) 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 prostate carcinoma tissue using anti-SPAK antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>, 1/50) for 30 minutes 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 prostate carcinoma tissue using anti-SPAK antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-37, 1/50) for 30 minutes 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 pancreas tissue using anti-SPAK antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7110-37" style="font-weight: bold;text-decoration: underline;">ET7110-37</a>, 1/50) for 30 minutes 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 pancreas tissue using anti-SPAK antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-37, 1/50) for 30 minutes 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.

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