Key Product Details

Species Reactivity

Validated:

Human, Rat

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Brevican
Asp23-Pro911
Accession # AAH27971

Specificity

Detects human Brevican in direct ELISAs and Western blots. In direct ELISAs, less than 20% cross-reactivity with recombinant mouse Brevican is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Rat Brevican Antibody

Detection of Human Brevican antibody by Western Blot.

Detection of Human Brevican by Western Blot.

Western blot shows lysates of human brain (cerebellum) tissue and human brain (motor cortex) tissue. PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human/Rat Brevican Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4009) followed by Donkey Anti-Sheep IgG HRP-conjugated Antigen Affinity-purified Polyclonal Antibody(Catalog # HAF016). Specific bands were detected for Brevican at approximately 60 and 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Brevican antibody in Rat Cortical Stem Cells by Immunocytochemistry (ICC).

Brevican in Rat Cortical Stem Cells.

Brevican was detected in immersion fixed rat cortical stem cells differentiated for 7 days by growth factor withdrawal using Sheep Anti-Human/Rat Brevican Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4009) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Brevican antibody in Human Brain by Immunohistochemistry (IHC-P).

Brevican in Human Brain.

Brevican was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Sheep Anti-Human/Rat Brevican Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4009) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to neuropil. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Brevican by Western Blot

Detection of Brevican by Western Blot

Brevican labeling in histological sections and western blots of human and rat prefrontal samples. (A) Micrographs and charts illustrating the average area fraction of brevican immunoreactivity in control and CUS exposed rats (four micrographs to the left and underlying charts) and in control and MDD human subjects (four micrographs to the right and underlying charts) showing no differences in overall brevican immunoreactivity and within 8 µm2 rectangles (see legends referring to rectangles in Figs. 2 And 3) around NRs both in animals with stress and in human subjects with MDD. Arrows point to nodes of Ranvier identified in the left two pictures using triple immunofluorescent staining for CASPR (labels paranodes in green) Neurofascin (red) and Brevican (blue in all the micrographs). Calibration bar is 5 µm for rat and 8 µm for human micrographs. (B) Representative western blot lanes and quantification of optical density of Brevican bands (relative to housekeeping protein actin) in the frontal pole of control and CUS rats, and in the PFC white matter of human subjects. The pictures present bands from 2 representative rats subjected to stress as compared to 2 controls (picture to the left) and two human MDD subjects against two non-psychiatric controls (picture to the right). Quantification for bands for all subjects in each group in the study at approximately 140 and 80 kD is presented in the charts. All lanes and bands in the pictures appear as they were positioned and developed in the Western blot PVDF membrane. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37775676), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Rat Brevican Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat cortical stem cells differentiated for 7 days by growth factor withdrawal

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cortex)

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Brevican (Catalog # 4009‑BC), see our available Western blot detection antibodies

Western Blot

0.1 µg/mL
Sample: Human brain (cerebellum) tissue and human brain (motor cortex) tissue

Reviewed Applications

Read 1 review rated 5 using AF4009 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Brevican

Brevican, also called BEHAB, is a secreted member of the the lectican family of proteoglycans that share a common domain structure (1). Brevican contains an Ig‑like V-set domain, two link domains, a Glu-rich region, a central region with glycosaminoglycan (GAG) modifications, an EGF-like domain, a C-type lectin domain, and a C-terminal Sushi/CRP-like domain (2). Brevican is restricted to the CNS and is expressed by astrocytes, oligodendrocytes, and neurons (3‑7). A GPI-anchored alternate splice form exists that is truncated following the central (GAG) region (2, 8). Brevican is cleaved by multiple proteases and exists in a number of distinct fragments (5, 9, 10). Full-length brevican consists of a 97 kDa core protein with up to approximately 100 kDa of attached chondroitin sulfate but not heparan sulfate chains (4, 7, 11, 12). Brevican associates with the extracellular matrix, perineuronal nets, and astrocyte cell surfaces by means of its chondroitin sulfate, GPI anchor, hyaluronic acid-binding link domains, and the core protein (4, 7, 8, 13). The secreted isoform is dominant during brain development and is up‑regulated in astrocytes following brain injury (2, 14). In human and rat, an under-glycosylated form of brevican is up‑regulated in highly aggressive glioma but not in low-grade glioma or other brain pathologies (15, 16). In mouse and rat, levels of an ADAMTS4-generated 55 kDa N-terminal fragment increase during remodeling after excitotoxic injury (11, 12). Human brevican shares 90%, 80%, and 80% aa sequence identity with bovine, mouse, and rat brevican, respectively. Within the Ig-like and two link domains, brevican shares 45%‑51% aa sequence identity with aggrecan, neurocan, and versican.

References

  1. Viapiano, M.S. and R.T. Matthews (2006) Trends Mol. Med. 12:488. 
  2. Gary, S.C. et al. (2000) Gene 256:139. 
  3. Jaworski, D.M. et al. (1994) J. Cell Biol. 125:495. 
  4. Seidenbecher, C.I. et al. (1995) J. Biol. Chem. 270:27206. 
  5. Hamel, M.G. et al. (2005) J. Neurochem. 93:1533. 
  6. Ogawa, T. et al. (2001) J. Comp. Neurol. 432:285  
  7. Yamada, H. et al. (1997) J. Neurosci. 17:7784.  
  8. Seidenbecher, C.I. et al. (2002) J. Neurochem. 83:738.
  9. Matthews, R.T. et al. (2000) J. Biol. Chem. 275:22695.
  10. Nakamura, H. et al. (2000) J. Biol. Chem. 275:38885.
  11. Mayer, J. et al. (2005) BMC Neurosci. 6:52.
  12. Yuan, W. et al. (2002) Neuroscience 114:1091.
  13. Deepa, S.S. et al. (2006) J. Biol. Chem. 281:17789.
  14. Jaworski, D.M. et al. (1999) Exp. Neurol. 157:327.
  15. Viapiano, M.S. et al. (2005) Cancer Res. 65:6726.
  16. Viapiano, M.S. et al. (2003) J. Biol. Chem. 278:33239.

Long Name

Brain Specific Proteoglycan in the Aggrecan Family

Alternate Names

ALPBRE, BCAN, BEHAB, CSPG7

Entrez Gene IDs

63827 (Human); 12032 (Mouse); 25393 (Rat)

Gene Symbol

BCAN

UniProt

Additional Brevican Products

Product Documents for Human/Rat Brevican Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Rat Brevican Antibody

For research use only

Citations for Human/Rat Brevican Antibody

Customer Reviews for Human/Rat Brevican Antibody (1)

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  • Human/Rat Brevican Antibody
    Name: Kimberly Alonge
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Adult brain
    Species: Rat
    Verified Customer | Posted 04/10/2018
    Used at a 1:500 but will need to increase to 1:100 for better staining
    Human/Rat Brevican Antibody AF4009

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