PRODUCT CODE: ET1612-20

Recombinant 44108 Monoclonal Antibody (ET1612-20)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • CHIP

REACTIVITY

  • Human

  • Mouse

Western blot analysis of Oct4 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 (ET1612-20, 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: F9 cell lysate<br />
Lane 2: HES cell lysate
  • Western blot analysis of Oct4 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 (ET1612-20, 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: F9 cell lysate<br />
Lane 2: HES cell lysate
  • ICC staining of Oct4 in N2A 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 (ET1612-20, 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 Oct4 in MCF-7 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 (ET1612-20, 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 mouse liver tissue using anti-Oct4 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 (ET1612-20, 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 Oct4 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 (ET1612-20, 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: F9 cell lysate
Lane 2: HES cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • CHIP

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant 44108 Monoclonal Antibody (ET1612-20)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

F9, NCCIT, hES, N2A, MCF-7, mouse liver tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD0750

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

39 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

POU domain, class 5, transcription factor 1

GENE NAME

POU5F1

SYNONYMS

Oct-4, Oct-3, OTF-3, POU5F1, OCT3, OCT4, OTF3

SEQUENCE SIMILARITIES

Belongs to the POU transcription factor family. Class-5 subfamily.

TISSUE SPECIFICITY

Expressed in developing brain. Highest levels found in specific cell layers of the cortex, the olfactory bulb, the hippocampus and the cerebellum. Low levels of expression in adult tissues.

DEVELOPMENTAL STAGE

Highly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation.

POST-TRANSLATIONAL MODIFICATION

Sumoylation enhances the protein stability, DNA binding and transactivation activity. Sumoylation is required for enhanced YES1 expression.; Ubiquitinated; undergoes 'Lys-63'-linked polyubiquitination by WWP2 leading to proteasomal degradation.; ERK1/2-mediated phosphorylation at Ser-111 promotes nuclear exclusion and proteasomal degradation. Phosphorylation at Thr-235 and Ser-236 decrease DNA-binding and alters ability to activate transcription.

SUBCELLULAR LOCATION

Cytoplasm. Nucleus. Note=Expressed in a diffuse and slightly punctuate pattern. Colocalizes with MAPK8 and MAPK9 in the nucleus.

FUNCTION

Transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). Forms a trimeric complex with SOX2 or SOX15 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. Critical for early embryogenesis and for embryonic stem cell pluripotency.

CITATIONS

  • Bin, Gu et al.

    Aire promotes the self-renewal of embryonic stem cells through Lin28. | Stem Cells and Development [2012]

  • Wang, Yongfang et al.

    N-myc downstream regulated gene 1(NDRG1) promotes the stem-like properties of lung cancer cells through stabilized c-Myc. | Cancer Letters [2017]