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STK33 Recombinant Rabbit Monoclonal Antibody [JE35-85]

Usd: 385

Specification

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Catalog# HA722395

STK33 Recombinant Rabbit Monoclonal Antibody [JE35-85]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

STK33 Recombinant Rabbit Monoclonal Antibody [JE35-85]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human STK33 aa 365-514 / 514.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC, IF-Tissue

Molecular Weight

Predicted band size: 58 kDa

Positive Control

Jurkat cell lysate, HEK-293 cell lysate, SK-Br-3 cell lysate, Jurkat, human testis tissue.

Conjugation

unconjugated

Clone Number

JE35-85

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:200

Target

Function

Serine/threonine protein kinase which phosphorylates VIME. May play a specific role in the dynamic behavior of the intermediate filament cytoskeleton by phosphorylation of VIME (By similarity). Not essential for the survival of KRAS-dependent AML cell lines.

Background References

1. Yu W et al. STK33 Phosphorylates Fibrous Sheath Protein AKAP3/4 to Regulate Sperm Flagella Assembly in Spermiogenesis. Mol Cell Proteomics. 2023 Jun

2. Röth S et al. Identification of KLHDC2 as an efficient proximity-induced degrader of K-RAS, STK33, beta-catenin, and FoxP3. Cell Chem Biol. 2023 Oct

Subcellular Location

Cytoplasm, perinuclear region.

UNIPROT

SwissProt: Q9BYT3 Human

Synonyms

Hypothetical protein FLJ35932 antibody

Serine threonine kinase 33 antibody

Serine/threonine kinase 33 antibody

Serine/threonine protein kinase 33 antibody

Serine/threonine-protein kinase 33 antibody

STK 33 antibody

Stk33 antibody

STK33_HUMAN antibody

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of STK33 on different lysates with Rabbit anti-STK33 antibody (<a href="/products/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</a>) at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: SK-Br-3 cell lysate (negative)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 58 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/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</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.

    ☑ Relative expression (RE)

    Western blot analysis of STK33 on different lysates with Rabbit anti-STK33 antibody (HA722395) at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: SK-Br-3 cell lysate (negative)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 58 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 (HA722395) 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.

  • Immunocytochemistry analysis of Jurkat cells labeling STK33 with Rabbit anti-STK33 antibody (<a href="/products/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-STK33 antibody (<a href="/products/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</a>) at 1/100 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 1/100 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 Jurkat cells labeling STK33 with Rabbit anti-STK33 antibody (HA722395) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-STK33 antibody (HA722395) at 1/100 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 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-STK33 antibody (<a href="/products/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</a>) at 1/1,000 dilution.<br /><br />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 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/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</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-STK33 antibody (HA722395) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722395) 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.

  • Flow cytometric analysis of Jurkat cells labeling STK33.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722395" style="font-weight: bold;text-decoration: underline;">HA722395</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 Jurkat cells labeling STK33.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722395, 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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