PRODUCT CODE: ET1612-99

14-3-3 alpha+beta Recombinant Rabbit Monoclonal Antibody [SD0837] (ET1612-99)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of 14-3-3 alpha+beta 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 (ET1612-99, 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: Hela cell lysate<br />
Lane 2: HepG2 cell lysate<br />
Lane 2: 293T cell lysate
  • Western blot analysis of 14-3-3 alpha+beta 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 (ET1612-99, 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: Hela cell lysate<br />
Lane 2: HepG2 cell lysate<br />
Lane 2: 293T cell lysate
  • ICC staining of 14-3-3 alpha+beta 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 (ET1612-99, 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 alpha+beta 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 (ET1612-99, 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 alpha+beta in SHG-44 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 (ET1612-99, 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).
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-14-3-3 alpha+beta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-99, 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 mouse brain tissue using anti-14-3-3 alpha+beta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-99, 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 mouse skin tissue using anti-14-3-3 alpha+beta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-99, 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 breast tissue using anti-14-3-3 alpha+beta antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-99, 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 14-3-3 alpha+beta was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-99, 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 alpha+beta 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 (ET1612-99, 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: Hela cell lysate
Lane 2: HepG2 cell lysate
Lane 2: 293T cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

14-3-3 alpha+beta Recombinant Rabbit Monoclonal Antibody [SD0837] (ET1612-99)

Immunogen

Synthetic peptide corresponding to c terminal human 14-3-3 alpha + beta aa 197-246 / 246.

Host

Rabbit

Positive Control

Hela cell lysate, HepG2 cell lysate, 293T cell lysate, Hela, A431, SHG-44, human breast carcinoma tissue, mouse brain tissue, mouse skin tissue, human breast tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD0837

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

  • 1:500-1:2,000

  • ICC

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

TARGET

UNIPROT #

PROTEIN NAME

14-3-3 alpha+beta

SYNONYMS

14 3 3 alpha antibody; 14 3 3 protein beta/alpha antibody; 14-3-3 protein beta/alpha antibody; 1433B_HUMAN antibody; Brain protein 14 3 3 beta isoform antibody; GW128 antibody; HS 1 antibody; KCIP-1 antibody; KCIP1 antibody; N-terminally processed antibody; Protein 1054 antibody; Protein kinase C inhibitor protein 1 antibody; YWHAA antibody; YWHAB antibody

SEQUENCE SIMILARITIES

Belongs to the 14-3-3 family.

POST-TRANSLATIONAL MODIFICATION

The alpha, brain-specific form differs from the beta form in being phosphorylated. Phosphorylated on Ser-60 by protein kinase C delta type catalytic subunit in a sphingosine-dependent fashion.

SUBCELLULAR LOCATION

Cytoplasm, Melanosome.

FUNCTION

Members of the 14-3-3 family of proteins are highly conserved proteins, localized in neurons, and are axonally transported to the nerve terminals. They are also present, at lower levels, in various other eukaryotic tissues. 14-3-3 proteins appear to play important roles in a variety of signal transduction pathways, including those involved in cell cycle regulation and cell survival. Because 14-3-3 proteins bind to specific phosphoserine-containing sequences they are likely to have an important role in signaling pathways mediated by serine/threonine protein kinases. Evidence indicates 14-3-3 is required for Raf 1 kinase activity and phosphorylation among many other functions.