Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab42453 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).Can be blocked with Human Olig2 peptide (ab111666). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
ICC/IF Use at an assay dependent concentration. PubMed: 20025521
IHC-Fr 1/200. (see Abreview and notes)
IHC (PFA fixed) 1/1000.

Target

  • Function
    Required for oligodendrocyte and motor neuron specification in the spinal cord, as well as for the development of somatic motor neurons in the hindbrain. Cooperates with OLIG1 to establish the pMN domain of the embryonic neural tube. Antagonist of V2 interneuron and of NKX2-2-induced V3 interneuron development.
  • Tissue specificity
    Expressed in the brain, in oligodendrocytes. Strongly expressed in oligodendrogliomas, while expression is weak to moderate in astrocytomas. Expression in glioblastomas highly variable.
  • Involvement in disease
    Note=A chromosomal aberration involving OLIG2 may be a cause of a form of T-cell acute lymphoblastic leukemia (T-ALL). Translocation t(14;21)(q11.2;q22) with TCRA.
  • Sequence similarities
    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Domain
    The bHLH is essential for interaction with NKX2-2.
  • Cellular localization
    Nucleus. Cytoplasm. The NLS contained in the bHLH domain could be masked in the native form and translocation to the nucleus could be mediated by interaction either with class E bHLH partner protein or with NKX2-2.
  • Information by UniProt
  • Database links
  • Alternative names
    • Basic domain helix loop helix protein class B 1 antibody
    • Basic helix loop helix protein class B 1 antibody
    • BHLHB antibody
    • bHLHB1 antibody
    • bHLHe19 antibody
    • Class B basic helix loop helix protein 1 antibody
    • Class B basic helix-loop-helix protein 1 antibody
    • class E basic helix loop helix protein 19 antibody
    • Class E basic helix-loop-helix protein 19 antibody
    • Human protein kinase C binding protein RACK17 antibody
    • Olig2 antibody
    • OLIG2_HUMAN antibody
    • Oligo2 antibody
    • Oligodendrocyte lineage transcription factor 2 antibody
    • Oligodendrocyte specific bHLH transcription factor 2 antibody
    • Oligodendrocyte transcription factor 2 antibody
    • OTTHUMP00000067569 antibody
    • OTTHUMP00000067570 antibody
    • PRKCBP2 antibody
    • Protein kinase C binding protein 2 antibody
    • Protein kinase C binding protein RACK17 antibody
    • Protein kinase C-binding protein 2 antibody
    • Protein kinase C-binding protein RACK17 antibody
    • RACK17 antibody
    see all

Images

  • Anti-Olig2 antibody (ab42453) at 0.5 µg/ml (in 5% skim milk/PBS buffer) + Jurkat cell lysate

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 32 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 28 kDa, 38 kDa. We are unsure as to the identity of these extra bands.

  • ab42453 at a 1/100 dilution staining Olig2 in human brain tissue (pontine base) by Immunohistochemistry (frozen sections) incubated for 1 hour at 20°C. Acetone fixed. Blocked with 10% milk for 30 minutes at 20°C. Secondary used at 1/200 pig polyclonal conjugated to biotin.

    Note nuclear and cytoplasmic staining of oligodendrocytes in the white matter and neurons in the grey matter.

    See Abreview

References

This product has been referenced in:
  • Alamir H  et al. In situ characterization of stem cells-like biomarkers in meningiomas. Cancer Cell Int 18:77 (2018). Read more (PubMed: 29849507) »
  • Sharma A  et al. Angiogenic Gene Signature Derived from Subtype Specific Cell Models Segregate Proneural and Mesenchymal Glioblastoma. Front Oncol 7:146 (2017). Read more (PubMed: 28744448) »
See all 4 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
BSA+NGS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (brain)
Specification
brain
Permeabilization
Yes - triton 0.2%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 19 2013

Answer

Gracias por contactarnos de nuevo.

Lamento mucho que sigáis teniendo problemas para detectar específicamente oligodendrocitos en vuestras muestras.

Revisando de nuevo el protocolo, y las imágenes enviadas, me gustaría hacer las siguientes sugerencias:

-Es posible que el proceso de sacrificado del animal y extracción del tejido haya podido provocar cierta degradación de éste. Si tenéis oportunidad, llevar a cabo fijación mediante perfusión del animal, para preservar mejor los tejidos.

Si necesitáis protocolos para llevar a cabo este proceso, tenemos en nuestra web disponibles que pueden resultar de utilidad:



https://www.abcam.com/index.html?pageconfig=resource&rid=11466
https://www.abcam.com/index.html?pageconfig=resource&rid=10188





-Dado que el proceso de fijación que habéis llevado a cabo en vuestro ensayo ha sido con 4% de formaldehido toda la noche, éste puede haber sido suficiente para crear puentes metilo y enmascarar los sitios antigénicos. Un método suave de recuperación antigénica puede mejorar la tinción. Yo recomendaría un método con calor (microondas u olla express) durante poco tiempo (máximo de 10 minutos), con un buffer tal que tEDTA o Citrato.

-Revisando las imágenes que nos enviaste en su día, me surgen algunas dudas. ¿ A que corresponden los distintos colores de las imágenes de fluorescencia? Según apuntan las flechas, el color rosa correspondería a los oligodendrocitos, y el amarillo y verde al resto de células. Intuyo que el azul corresponde a los núcleos teñidos con DAPI, pero, ¿Cómo se han obtenido los colores amarillo, verde? ¿Cómo se ha comprobado que esa tinción corresponde a los axones u astrocitos? ¿Se ha usado algún marcador de axones? Si no es así, quizás esta pudiera ser una posible acción a tomar.

-¿Cómo se han llevado a cabo los cortes de las secciones? ¿longitudinal o transversalmente? ¿Qué zona de la medula espinal se usó, craneal, dorsal, sacra, o lumbar?

-Como ultima sugerencia se me ocurre que quizás queráis probar, como control positivo, para evaluar el rendimiento del anticuerpo, unas condiciones similares a las que vienen en la datasheet. El anticuerpo ab81093 lo hemos testado en nuestro laboratorio usando “human normal cerebral cortex formalin fixed paraffin embedded tissue section”.

Podríamos mandaros (sin cargos) una seccione del catalogo similar a la que se usó con el anticuerpo, y tratáis de reproducir las condiciones, y así salir de dudas respecto al origen de la inespecificidad de la tinción. Las secciones que podrían valer para ello son:



https://www.abcam.com/Brain-human-frontal-lobe-normal-tissue-slides-ab4304.html
https://www.abcam.com/Brain-human-parietal-lobe-normal-tissue-slides-ab4313.html



Espero que alguna de estas ideas pueda seros de ayuda. Espero tu respuesta y me confirmas cuál de ellas quisierais probar.

Que paséis un buen día.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (E10.5 mouse embryo sections)
Specification
E10.5 mouse embryo sections
Fixative
Paraformaldehyde
Permeabilization
Yes - PBST (PBS/0.5%v/v TritonX100 (Sigma)
Blocking step
(agent) for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 08 2012

Answer

Thank you very much for contacting us.

If you could specify which image you are referring to, I can try to help you further. However, regarding the references, I am sorry to let you know that we do not have unrestricted access to all journals. So if the reference cited on the datasheet is not freely accessible, we will not have it either and thus cannot pass it on to you.

Having said this, I am very happy to find out for you where the image comes from. Quite a few of our images originate from other users who submitted an Abreview about their results with the product. You can access these Abreviews in the respective section on each datasheet. Often the researchers agreed to be contacted by others (unfortunately not by us), so you might find further details on each Abreview page.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Brain - pontine base)
Specification
Brain - pontine base
Fixative
Acetone
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 15 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain - frontal cortex)
Specification
Brain - frontal cortex
Fixative
Formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer (pH6), EDTA (pH8) and dionized water
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Sep 23 2009

Answer

Thank you for your patience in this matter. I have received a reply from the originator of ab42453 (Olig2 antibody). The exact immunogenic peptide sequence is proprietary information. However, we can state that it is within the following 50 amino acid sequence: SLPGSGLPSVGSIRPPHGLLKSPSAAAAAPLGGGGGGSGASGGFQHWGGM I hope this information helps. Please don't hesitate to contact us again if you have further questions.

Read More

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