Recombinant

Recombinant Anti-Myelin Basic Protein antibody [IGX3421] (ab209328)

Human recombinant monoclonal Myelin Basic Protein antibody [IGX3421]. Validated in WB, ELISA, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 1 publication(s). Independently reviewed in 3 review(s).

Overview

  • Product name

    Anti-Myelin Basic Protein antibody [IGX3421]
    See all Myelin Basic Protein primary antibodies
  • Description

    Human monoclonal [IGX3421] to Myelin Basic Protein
  • Host species

    Human
  • Tested applications

    Suitable for: ELISA, WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Cow, Cat, Dog, Pig, Chimpanzee, Macaque monkey
  • Immunogen

    Full length native protein (purified). This information is considered to be commercially sensitive.
    (Peptide available as ab792)

  • Positive control

    • WB: Human, mouse and rat brain tissue lysate. Myelin Basic Protein (recombinant protein) IHC-P: Mouse, rat and human brain tissue (Hippocampus) ICC/IF: SK-N-SH cells
  • General notes

    This product was made using synthetic libraries and phage display technology.

    This antibody is a recombinant antibody.  
    Human monoclonal antibody.

     

    Example of usage (reference):

    Spatiotemporal Dynamics of Molecular Pathology in Amyotrophic Lateral Sclerosis

    Silas Maniatis, Tarmo Aijo, Sanja Vickovic, Catherine Braine, Kristy Kang, Annelie Mollbrink, Zaneta Andrusivova, Sami Saarenpaa, Gonzalo Saiz-Castro, Miguel Cuevas, Aaron Watters, Joakim Lundeberg, Richard Bonneau, Hemali Phatnani

Properties

Applications

Our Abpromise guarantee covers the use of ab209328 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
ELISA Use at an assay dependent concentration.
WB Use a concentration of 0.25 - 1 µg/ml. Detects a band of approximately 20,17 kDa (predicted molecular weight: 33 kDa).
ICC/IF Use a concentration of 5 µg/ml.

This product gave a positive signal in SKNSH cells fixed with 4% formaldehyde

IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    The classic group of MBP isoforms (isoform 4-isoform 14) are with PLP the most abundant protein components of the myelin membrane in the CNS. They have a role in both its formation and stabilization. The smaller isoforms might have an important role in remyelination of denuded axons in multiple sclerosis. The non-classic group of MBP isoforms (isoform 1-isoform 3/Golli-MBPs) may preferentially have a role in the early developing brain long before myelination, maybe as components of transcriptional complexes, and may also be involved in signaling pathways in T-cells and neural cells. Differential splicing events combined with optional post-translational modifications give a wide spectrum of isomers, with each of them potentially having a specialized function. Induces T-cell proliferation.
  • Tissue specificity

    MBP isoforms are found in both the central and the peripheral nervous system, whereas Golli-MBP isoforms are expressed in fetal thymus, spleen and spinal cord, as well as in cell lines derived from the immune system.
  • Involvement in disease

    Note=The reduction in the surface charge of citrullinated and/or methylated MBP could result in a weakened attachment to the myelin membrane. This mechanism could be operative in demyelinating diseases such as chronical multiple sclerosis (MS), and fulminating MS (Marburg disease).
  • Sequence similarities

    Belongs to the myelin basic protein family.
  • Developmental stage

    Expression begins abruptly in 14-16 week old fetuses. Even smaller isoforms seem to be produced during embryogenesis; some of these persisting in the adult. Isoform 4 expression is more evident at 16 weeks and its relative proportion declines thereafter.
  • Post-translational
    modifications

    Several charge isomers of MBP; C1 (the most cationic, least modified, and most abundant form), C2, C3, C4, C5, C6, C7, C8-A and C8-B (the least cationic form); are produced as a result of optional PTM, such as phosphorylation, deamidation of glutamine or asparagine, arginine citrullination and methylation. C8-A and C8-B contain each two mass isoforms termed C8-A(H), C8-A(L), C8-B(H) and C8-B(L), (H) standing for higher and (L) for lower molecular weight. C3, C4 and C5 are phosphorylated. The ratio of methylated arginine residues decreases during aging, making the protein more cationic.
    The N-terminal alanine is acetylated (isoform 3, isoform 4, isoform 5 and isoform 6).
    Arg-241 was found to be 6% monomethylated and 60% symmetrically dimethylated.
  • Cellular localization

    Myelin membrane. Cytoplasmic side of myelin.
  • Information by UniProt
  • Database links

  • Alternative names

    • GDB antibody
    • Golli MBP antibody
    • Golli MBP; myelin basic protein antibody
    • Hemopoietic MBP antibody
    • HMBPR antibody
    • HUGO antibody
    • MBP antibody
    • MBP_CAVPO antibody
    • MBP_HUMAN antibody
    • MGC99675 antibody
    • MLD antibody
    • Myelin A1 protein antibody
    • Myelin A1 Protein, basic antibody
    • Myelin basic protein antibody
    • Myelin Deficient antibody
    • Myelin membrane encephalitogenic protein antibody
    • OTTHUMP00000163776 antibody
    • OTTHUMP00000174387 antibody
    • OTTHUMP00000174388 antibody
    • SHI antibody
    • Shiverer antibody
    • SP antibody
    see all

Images

  • IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal rat brain and normal rat pancreas, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

    An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab209328 staining Myelin Basic Protein in SK-N-SH (Human neuroblastoma cell line) cells. The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal donkey serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab209328 at a 5 µg/ml concentration, then detected with a donkey anti-human (Alexa Fluor® 488) secondary antibody at a 1/2000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal human hippocampus and normal human pancreas*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

    An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

     *Human pancreas tissue was obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC image of Myelin Basic Protein staining in a section of formalin-fixed paraffin-embedded normal mouse brain and normal mouse pancreas, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab209328, 1/1000 dilution, for 15 minutes at room temperature.

    An HRP-conjugated goat anti-Human IgG secondary was used for 15 minutes at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Myelin Basic Protein antibody [IGX3421] (ab209328) at 0.25 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466) at 10 µg
    Lane 2 : Mouse brain tissue lysate at 10 µg
    Lane 3 : Rat brain tissue lysate at 10 µg
    Lane 4 : Myelin Basic Protein (Recombinant protein) at 0.1 µg

    Secondary
    All lanes : HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 18,23,24 kDa
    why is the actual band size different from the predicted?



    Exposure time :

    Lane 1 : 30 seconds.

    Lanes 2-3 : 2 minutes.

    Lane 4 : 8 minutes.

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209328 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • ELISA using ab209328 for 16 hours at 4ºC. ab7153 goat anti human was used as a secondary at a 1/5000 dilution for 1 hour at Room Temperature.

  • Anti-Myelin Basic Protein antibody [IGX3421] (ab209328) at 1 µg/ml + Mouse brain tissue lysate at 10 µg

    Secondary
    HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 20, 17 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209328 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

This product has been referenced in:

  • Chen Z  et al. Hyperglycemia aggravates spinal cord injury through endoplasmic reticulum stress mediated neuronal apoptosis, gliosis and activation. Biomed Pharmacother 112:108672 (2019). Read more (PubMed: 30784940) »
  • Witt MR  et al. Calcium homeostasis in fibroblasts from patients with amyotrophic lateral sclerosis. J Neurol Sci 126:206-12 (1994). Read more (PubMed: 7853028) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 2°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 23 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 23 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Peripheral nerve)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Peripheral nerve
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 23 2016

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