PRODUCT CODE: ET1612-97

Recombinant 14-3-3 Theta Monoclonal Antibody (ET1612-97)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of 14-3-3 Theta 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 (ET1612-97, 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: MCF-7 cell lysate<br />
 Lane 2: Hela cell lysate
  • Western blot analysis of 14-3-3 Theta 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 (ET1612-97, 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: MCF-7 cell lysate<br />
 Lane 2: Hela cell lysate
  • 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 (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 (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 (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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of 14-3-3 Theta 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 (ET1612-97, 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: MCF-7 cell lysate
Lane 2: Hela cell lysate

Applications

  • WB

  • ICC

  • IF

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant 14-3-3 Theta Monoclonal Antibody (ET1612-97)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

MCF-7 cell lysate, Hela cell lysate, Hela, A431, A549, SH-SY5Y.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD084-02

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. 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

28 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • FC

  • 1:50-1:100

  • ICC/IF

  • 1:100-1:500

TARGET

UNIPROT #

PROTEIN NAME

14-3-3 Theta

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

SEQUENCE SIMILARITIES

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.

FUNCTION

14-3-3 proteins regulate many cellular processes relevant to cancer biology, notably apoptosis, mitogenic signaling and cell-cycle checkpoints. Seven isoforms comprise this family of signaling intermediates, denoted 14-3-3 β, γ, ε, ζ, η, θ and σ. 14-3-3 proteins form dimers that present two binding sites for ligand proteins, thereby bringing together two proteins that may not otherwise associate. These ligands largely share a 14-3-3 consensus binding motif and exhibit serine/threonine phosphorylation. 14-3-3 proteins function in broad regulation of these ligand proteins, by cytoplasmic sequestration, occupation of interaction domains and import/export sequences, prevention of degradation, activation/repression of enzymatic activity and facilitation of protein modification, and thus loss of expression contributes to a vast array of pathogenic cellular activities.