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Oct4 Mouse Monoclonal Antibody [A3-C11]

Usd: 350

Specification

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Catalog# EM100306

Oct4 Mouse Monoclonal Antibody [A3-C11]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Oct4 Mouse Monoclonal Antibody [A3-C11]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within full length human Oct4 protein.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 39 kDa

Positive Control

NCCIT cell lysate, D3 cell lysate, NCCIT.

Conjugation

unconjugated

Clone Number

A3-C11

RRID

AB_3068737

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*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:200

  • FC

  • 1:1000

Target

Function

Oct-4 (POU5F1) is transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). It forms a trimeric complex with SOX2 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. Oct-4 is critical for early embryogenesis and for embryonic stem cell pluripotency. It is expressed in developing brain and highest levels also found in specific cell layers of the cortex, the olfactory bulb, the hippocampus and the cerebellum.

Background References

1. Oct4 pseudogenes are transcribed in cancers."Suo G., Han J., Wang X., Zhang J., Zhao Y., Zhao Y., Dai J.Biochem. Biophys. Res. Commun. 337:1047-1051(2005)

2. Sumoylation of Oct4 enhances its stability, DNA binding, and transactivation."Wei F., Schoeler H.R., Atchison M.L.J. Biol. Chem. 282:21551-21560(2007)

3. Post-translational modification of POU domain transcription factor Oct-4 by SUMO-1."Zhang Z., Liao B., Xu M., Jin Y.FASEB J. 21:3042-3051(2007)

Sequence Similarity

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.

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.

UNIPROT

SwissProt: Q01860 Human

Synonyms

Octamer binding transcription factor 4 antibody

MGC22487 antibody

Oct 3 antibody

Oct 4 antibody

Oct-3 antibody

Oct-4 antibody

OCT3 antibody

Oct4 antibody

Octamer binding protein 3 antibody

Octamer binding protein 4 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Oct4 on different lysates with Mouse anti-Oct4 antibody (<a href="/products/EM100306" style="font-weight: bold;text-decoration: underline;">EM100306</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Oct4 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 39 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 6 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/EM100306" style="font-weight: bold;text-decoration: underline;">EM100306</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Oct4 on different lysates with Mouse anti-Oct4 antibody (EM100306) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Oct4 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 39 kDa
    Observed band size: 42 kDa

    Exposure time: 6 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 (EM100306) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Oct4 on different lysates using anti-Oct4 antibody at 1/1,000 dilution.<br /><span style="font-weight: bold;">Positive control:</span><br />Lane 1: NCCIT cell lysate<br />Lane 2: D3 cell lysate

    Western blot analysis of Oct4 on different lysates using anti-Oct4 antibody at 1/1,000 dilution.
    Positive control:
    Lane 1: NCCIT cell lysate
    Lane 2: D3 cell lysate

  • ICC staining of OCT-4 in NCCIT cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining of OCT-4 in NCCIT cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Flow cytometric analysis of NCCIT cells labeling Oct4.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/EM100306" style="font-weight: bold;text-decoration: underline;">EM100306</a>, 1:1000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NCCIT cells labeling Oct4.

    Cells were fixed and permeabilized. Then stained with the primary antibody (EM100306, 1:1000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation