PRODUCT CODE: ET7110-37

SPAK Recombinant Rabbit Monoclonal Antibody [JE53-64] (ET7110-37)

  • Recombinant

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

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Western blot analysis of SPAK on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: HepG2 cell lysate
  • Western blot analysis of SPAK on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: HepG2 cell lysate
  • 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) for 20 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) for 20 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.
  • Flow cytometric analysis of SPAK was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-37, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of SPAK on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Jurkat cell lysate
Lane 2: HepG2 cell lysate

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

SPAK Recombinant Rabbit Monoclonal Antibody [JE53-64] (ET7110-37)

Immunogen

Synthetic peptide corresponding to c terminal human spak.

Host

Rabbit

Positive Control

Jurkat cell lysate, HepG2 cell lysate, human prostate carcinoma tissue, human pancreas tissue, Daudi.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JE53-64

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

59 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

SPAK

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; SPAK antibody; Ste 20 related kinase antibody; Ste-20-related kinase antibody; Ste20 like protein kinase antibody; STE20/SPS1 homolog antibody; STE20/SPS1 related proline alanine rich protein kinase antibody; STE20/SPS1-related proline-alanine-rich protein kinase antibody; STK 39 antibody; Stk39 antibody; STK39_HUMAN antibody

SEQUENCE SIMILARITIES

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

Nucleus, cytoplasm.

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.