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14-3-3 Theta Recombinant Rabbit Monoclonal Antibody [SD084-02]

Usd: 385

Specification

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Catalog# ET1612-97

14-3-3 Theta Recombinant Rabbit Monoclonal Antibody [SD084-02]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • FC

Reactivity

  • Human

  • Rat

Predicted reactivity

  • Mouse

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

14-3-3 Theta Recombinant Rabbit Monoclonal Antibody [SD084-02]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human 14-3-3 Theta aa 1-50 / 245.

Species Reactivity

Human, Rat (Predicted: Mouse)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, FC

Molecular Weight

Predicted band size: 28 kDa

Positive Control

Rat testis tissue lysate, rat brain tissue lysate, C6 cell lysate, MCF7 cell lysate, Hela, A431, A549, SH-SY5Y.

Conjugation

unconjugated

Clone Number

SD084-02

RRID

AB_3070173

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

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

  • 1:500-1:5,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:100-1:500

  • FC

  • 1:50-1:100

Target

Function

14-3-3 protein theta is a protein that in humans is encoded by the YWHAQ gene. This gene product belongs to the 14-3-3 family of proteins that mediate signal transduction by binding to phosphoserine-containing proteins. This highly conserved protein family is found in both plants and mammals, and this protein is 99% identical to the mouse and rat orthologs. This gene is upregulated in patients with amyotrophic lateral sclerosis. It contains in its 5' UTR a 6 bp tandem repeat sequence that is polymorphic; however, there is no correlation between the repeat number and the disease.

Background References

1. Scheibner, KA. et al. 2012. MiR-27a functions as a tumor suppressor in acute leukemia by regulating 14-3-3θ. PLoS ONE. 7: e50895.

2. Karasawa, T. et al. 2010. CLIMP-63 is a gentamicin-binding protein that is involved in drug-induced cytotoxicity. Cell death & disease. 1: e102.

Sequence Similarity

Belongs to the 14-3-3 family.

Tissue Specificity

Abundantly expressed in brain, heart and pancreas, and at lower levels in kidney and placenta. Up-regulated in the lumbar spinal cord from patients with sporadic amyotrophic lateral sclerosis (ALS) compared with controls, with highest levels of expression in individuals with predominant lower motor neuron involvement.

Post-translational Modification

Ser-232 is probably phosphorylated by CK1.

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: P27348 Human

SwissProt: P68254 Mouse

SwissProt: P68255 Rat

Synonyms

14-3-3 antibody

14-3-3 protein T-cell antibody

14-3-3 protein tau antibody

14-3-3 protein theta antibody

1C5 antibody

HS1 antibody

theta polypeptide antibody

tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein antibody

tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, theta isoform antibody

tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, theta polypeptide antibody

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of 14-3-3 Theta on different lysates with Rabbit anti-14-3-3 Theta antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-14-3-3 Theta KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 28 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 46 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/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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.

    ☑ Knockdown (KD)

    Western blot analysis of 14-3-3 Theta on different lysates with Rabbit anti-14-3-3 Theta antibody (ET1612-97) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-14-3-3 Theta KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 28 kDa
    Observed band size: 28 kDa

    Exposure time: 46 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 (ET1612-97) at 1/1,000 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.

  • Western blot analysis of 14-3-3 Theta on different lysates with Rabbit anti-14-3-3 Theta antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>) at 1/1,000 dilution.<br /><br />Lane 1: Rat testis tissue lysate<br />Lane 2: Rat brain tissue lysate<br />Lane 3: C6 cell lysate<br />Lane 4: MCF7 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 28 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 10 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/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</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.

    Western blot analysis of 14-3-3 Theta on different lysates with Rabbit anti-14-3-3 Theta antibody (ET1612-97) at 1/1,000 dilution.

    Lane 1: Rat testis tissue lysate
    Lane 2: Rat brain tissue lysate
    Lane 3: C6 cell lysate
    Lane 4: MCF7 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 28 kDa
    Observed band size: 28 kDa

    Exposure time: 10 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 (ET1612-97) 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.

  • ICC staining of 14-3-3 Theta in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of 14-3-3 Theta in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of 14-3-3 Theta in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of 14-3-3 Theta in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of 14-3-3 Theta in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of 14-3-3 Theta in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Flow cytometric analysis of 14-3-3 Theta was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1612-97" style="font-weight: bold;text-decoration: underline;">ET1612-97</a>, 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of 14-3-3 Theta was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-97, 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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