The complexity of human astrocytes stays poorly outlined in major human tissue, requiring higher instruments for his or her isolation and molecular characterization. Fluorescence-activated nuclei sorting (FANS) can be utilized to efficiently isolate and examine human neuronal nuclei (NeuN+) populations from frozen archival tissue, thereby avoiding issues related to dealing with recent tissue. Nonetheless, efforts to equally isolate astroglia from the non-neuronal (NeuN-) ingredient are missing.
A lately developed and validated immunotagging technique makes use of three transcription issue antibodies to concurrently isolate enriched neuronal (NeuN+), astrocyte (paired field protein 6 (PAX6)+NeuN-), and oligodendrocyte progenitor (OLIG2+NeuN-) nuclei populations from non-diseased, recent (unfixed) snap-frozen postmortem human temporal neocortex tissue.
This system was proven to be helpful for the characterization of cell type-specific transcriptome alterations in major pathological epilepsy neocortex. Transcriptomic analyses confirmed that PAX6+NeuN- sorted populations are robustly enriched for pan-astrocyte markers and seize astrocytes in each resting and reactive circumstances.
This paper describes the FANS methodology for the isolation of astrocyte-enriched nuclei populations from fresh-frozen human cortex, together with tissue dissociation into single-nucleus (sn) suspension; immunotagging of nuclei with anti-NeuN and anti-PAX6 fluorescently conjugated antibodies; FANS gating methods and high quality management metrics for optimizing sensitivity and specificity throughout sorting and for confirming astrocyte enrichment; and really helpful procurement for downstream transcriptome and chromatin accessibility sequencing at bulk or sn decision. This protocol is relevant for non-necrotic, fresh-frozen, human cortical specimens with numerous pathologies and really helpful postmortem tissue assortment inside 24 h.

Principal Function of Antibodies in Demyelination and Axonal Harm in A number of Sclerosis

Antibodies and oxidative stress are hallmarks of a number of sclerosis (MS) lesions. We aimed to make clear the relation between them, their position in MS sufferers and to research their specificity, evaluating MS with classical neurodegenerative ailments (ND). Mind samples from 14 MS instances, 6 with ND and 9 controls (with out neurological ailments). Immunohistochemistry assays have been used to detect oxidized lipids (EO6), IgG and IgM, oligodendrocytes (Olig2), axons (NF, neurofilament) and mobile (TUNEL) and axonal harm (APP, amyloid precursor protein).
We didn’t observe EO6 in controls. All samples from MS sufferers confirmed EO6 in oligodendrocytes and axons inside lesions. We didn’t detect co-localization between EO6 and antibodies. Neither did we between EO6 and TUNEL or APP. 94.4% of TUNEL-positive cells in regular showing white matter have been additionally stained for IgG and 75.5% for IgM. IgM, however not IgG, co-localized with APP. EO6 was related to axonal harm in amyotrophic lateral sclerosis (ALS).
We didn’t observe affiliation between antibodies and mobile or axonal harm in ND sufferers. MS sufferers confirmed a better variety of B cells and plasma cells within the lesions and meninges than controls. The variety of B cells and plasma cells was related to the presence of antibodies and with the exercise of the lesions. We noticed a predominant position of B lymphocytes within the improvement of MS lesions. Antibodies contribute to the oligodendrocyte and axonal harm in MS. Oxidative stress was related to axonal harm in ALS.

Correlation Between Immunohistochemistry and Sequencing in H3G34-Mutant Gliomas

Recurrent glycine-to-arginine/valine alterations at codon 34 (G34R/V) inside H3F3A gene characterize a subset of hemispheric high-grade gliomas (HGG) affecting kids and younger adults. These tumors, outlined as G34R/V-mutant gliomas, are histologically heterogenous, with microscopic options of both HGG or embryonal tumors (primitve neuroectodermal tumor-like options).
To evaluate the worth of immunohistochemistry (IHC) to detect G34R/V-mutated instances, we examined anti-histone G34V (clone 329E5) and anti-histone G34R (clone RM240) antibodies in a sequence of 28 formalin-fixed and paraffin-embedded samples. A complete of 28 instances of hemispheric, IDH-wt HGG primarily affecting kids and younger adults have been evaluated by IHC and by sequencing. The median age of sufferers at analysis was 17 years (0.1 to 26 y). By IHC, 10 of the 28 instances confirmed nuclear positivity for G34R and three of the 28 instances for G34V.
Molecular evaluation of G34R/V-mutation standing was profitable in 24 of the 28 instances. Mutation at glycine 34 of the H3F3A gene was recognized in 9 of the 24 tumors (37%) by direct sequencing, revealing 7 of 9 constructive case by sequencing and a pair of of 9 false destructive instances by IHC.
Two of 15 destructive case by sequencing demonstrated a false positivity by IHC. In whole, in 4 (16.6%) of 24 instances, IHC and mutational outcomes have been discordant: 2 tumors have been destructive by IHC (false destructive) however harbored G34R mutation by sequencing, and a pair of instances have been constructive by IHC (false constructive by IHC) however wild kind by sequencing.
Furthermore, most mutated instances confirmed lack of ATRX expression and/or p53 expression. The positivity by IHC with particular antibody examined just isn’t extremely predictive for presence of G34R/V mutation, however affirmation by sequencing is necessary; G34R/V mutations ought to be suspected in all hemispheric tumor IDH1/2 wild kind, exhibiting lack of OLIG2 and ATRX and/or p53 expression.

Oligodendrocyte Lineage Marker Expression in eGFP-GFAP Transgenic Mice

Oligodendrocytes, the myelinating cells of the central nervous system, orchestrate a number of key mobile capabilities within the mind and spinal wire, together with axon insulation, power switch to neurons, and, ultimately, modulation of immune responses. There’s rising curiosity for acquiring dependable markers that may particularly label oligodendroglia and their progeny.
In lots of research, anti-CC1 antibodies, presumably recognizing the protein adenomatous polyposis coli (APC), are used to label mature, myelinating oligodendrocytes. Nonetheless, it has been mentioned whether or not anti-CC1 antibodies may acknowledge as effectively, beneath pathological circumstances, different cell populations, significantly astrocytes.

Olig2 Antibody

F46745-0.08ML 0.08 ml
EUR 140.25
Description: This gene encodes a basic helix-loop-helix transcription factor which is expressed in oligodendroglial tumors of the brain. The protein is an essential regulator of ventral neuroectodermal progenitor cell fate. The gene is involved in a chromosomal translocation t(14;21)(q11.2;q22) associated with T-cell acute lymphoblastic leukemia. Its chromosomal location is within a region of chromosome 21 which has been suggested to play a role in learning deficits associated with Down syndrome. [provided by RefSeq].

Olig2 Antibody

F46745-0.4ML 0.4 ml
EUR 322.15
Description: This gene encodes a basic helix-loop-helix transcription factor which is expressed in oligodendroglial tumors of the brain. The protein is an essential regulator of ventral neuroectodermal progenitor cell fate. The gene is involved in a chromosomal translocation t(14;21)(q11.2;q22) associated with T-cell acute lymphoblastic leukemia. Its chromosomal location is within a region of chromosome 21 which has been suggested to play a role in learning deficits associated with Down syndrome. [provided by RefSeq].

OLIG2 Antibody

DF8004 200ul
EUR 420

OLIG2 Antibody

DF8004-100ul 100ul
EUR 280

OLIG2 Antibody

DF8004-200ul 200ul
EUR 350

OLIG2 antibody

E39-05987 100ug/100ul
EUR 225
Description: Available in various conjugation types.

OLIG2 Antibody

45197-100ul 100ul
EUR 302.4

OLIG2 Antibody

45197-50ul 50ul
EUR 224.4

OLIG2 Antibody

7743-002mg 0.02 mg
EUR 206.18
Description: The oligodendrocyte transcription factors 1 and 2 (OLIG1 and OLIG2, respectively) make up part of basic helix-loop-helix (bHLH) family of transcription factors that are specifically expressed in zones of the neuroepithelium from which oligodendrocyte precursors emerge (1). Both OLIG1 and OLIG2 genes are downstream targets of Sonic hedgehog and are expressed exclusively in the central nervous system (2). OLIG2 is first observed in the ventral most p3 progenitor domain of the ventral neural tube while OLIG1 is first expressed in the dorsal portion of the p3 domain (2). Mice overexpressing OLIG2 exhibit impaired potassium channel expression in neural progenitors and proliferation of these cells similar to that seen in Down Syndrome, suggesting that OLIG2 may play a role in this pathology (3).

OLIG2 Antibody

7743-01mg 0.1 mg
EUR 523.7
Description: The oligodendrocyte transcription factors 1 and 2 (OLIG1 and OLIG2, respectively) make up part of basic helix-loop-helix (bHLH) family of transcription factors that are specifically expressed in zones of the neuroepithelium from which oligodendrocyte precursors emerge (1). Both OLIG1 and OLIG2 genes are downstream targets of Sonic hedgehog and are expressed exclusively in the central nervous system (2). OLIG2 is first observed in the ventral most p3 progenitor domain of the ventral neural tube while OLIG1 is first expressed in the dorsal portion of the p3 domain (2). Mice overexpressing OLIG2 exhibit impaired potassium channel expression in neural progenitors and proliferation of these cells similar to that seen in Down Syndrome, suggesting that OLIG2 may play a role in this pathology (3).

OLIG2 antibody

70R-15037 100 ul
EUR 497
Description: Rabbit polyclonal OLIG2 antibody

OLIG2 antibody

70R-19040 50 ul
EUR 289
Description: Rabbit polyclonal OLIG2 antibody

OLIG2 Antibody

ABD8004 100 ug
EUR 525.6

olig2 Antibody

CAC11579-100ug 100ug
EUR 314

olig2 Antibody

CAC11579-50ug 50ug
EUR 199.2

OLIG2 antibody

CAF50242-100ug 100ug
EUR 338

olig2 Antibody

1-CSB-PA24549A0Rb
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  • Ask for price
  • 100ug
  • 50ug
Description: A polyclonal antibody against olig2. Recognizes olig2 from Zebrafish. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:5000

OLIG2 Antibody

GWB-BAACC1 0.05 mg Ask for price
On this examine, we used transgenic mice by which astrocytes are labeled by the improved inexperienced fluorescent protein (eGFP) beneath the management of the human glial fibrillary acidic protein (GFAP) promoter. By detailed co-localization research we have been in a position to reveal {that a} important proportion of eGFP-expressing cells co-express markers of the oligodendrocyte lineage, such because the transcription issue Oligodendrocyte Transcription Issue 2 (OLIG2); the NG2 proteoglycan, also referred to as chrondroitin sulfate proteoglycan 4 (CSPG4); or APC. The present discovering that the GFAP promoter drives transgene expression in cells of the oligodendrocyte lineage ought to be thought-about when deciphering outcomes from co-localization research.

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