PRODUCT CODE: ET1701-2

14-3-3 Recombinant Rabbit Monoclonal Antibody [JJ083-6] (ET1701-2)

  • Recombinant

Applications

  • WB

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of 14-3-3 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 (ET1701-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Human tonsil tissue lysate<br />
Lane 2: HT-29 cell lysate<br />
Lane 3: NIH/3T3 cell lysate
  • Western blot analysis of 14-3-3 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 (ET1701-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Human tonsil tissue lysate<br />
Lane 2: HT-29 cell lysate<br />
Lane 3: NIH/3T3 cell lysate
  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-14-3-3 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 (ET1701-2, 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.
Western blot analysis of 14-3-3 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 (ET1701-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Human tonsil tissue lysate
Lane 2: HT-29 cell lysate
Lane 3: NIH/3T3 cell lysate

Applications

  • WB

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

14-3-3 Recombinant Rabbit Monoclonal Antibody [JJ083-6] (ET1701-2)

Immunogen

Recombinant protein within human 14-3-3 aa 1-126 / 246.

Host

Rabbit

Positive Control

NIH/3T3, HT29, human skin tissue, mouse skin tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ083-6

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 purified.

MOLECULAR WEIGHT

28 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

14-3-3

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

The 14-3-3 family of proteins are important regulatory molecules ubiquitiously expressed in all eukarryotic cells which bind to numerous signaling proteins in various pathways driving critical cellular pathways of apoptosis, differentiation and cell cycle. 14-3-3 theta specifically has also been shown to play an important regulatory role in the TLR2 signaling pathways as a negative regulator of TLR2 ligand Pam3CySk4 induced NF-kB activation. 14-3-3 theta has previously been shown to interact with TLR4 ligand and MyD88 dependent phosphorylated PkC epsilon. 14-3-3 theta in the TLR4 signaling pathway is a positive regulator controlling release of IRF3 indcuded pro-inflammatory cytokines RANTES and IP-10. Currently identified by mass spec as part of the TLR2 signaling complex and taken along with TLR4 data, a 14-3-3 theta antibody can be used to examine the different regulatory functions of 14-3-3 theta for different TLRs through its interaction with common or unique TLR signaling adaptor molecules in addition to MyD88 or PkC epsilon such as TRAM or TRIF allowing further clarification of TLR specific pathway regulation.