Recombinant Anti-Myelin Basic Protein antibody [IGX3421] (ab209328)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Human monoclonal [IGX3421] to Myelin Basic Protein
- Suitable for: ELISA, WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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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, Recombinant fragment
Predicted to work with: Horse, Cow, Cat, Dog, Pig, Chimpanzee, Macaque monkey -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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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
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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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
IGX3421 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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ELISA |
Use at an assay dependent concentration.
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WB | (2) |
Use a concentration of 0.25 - 1 µg/ml. Detects a band of approximately 20,17 kDa (predicted molecular weight: 33 kDa).
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ICC/IF |
Use a concentration of 5 µg/ml.
This product gave a positive signal in SKNSH cells fixed with 4% formaldehyde |
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IHC-P | (6) |
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.
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Notes |
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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
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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
modificationsSeveral 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
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Database links
- Entrez Gene: 618684 Cow
- Entrez Gene: 4155 Human
- Entrez Gene: 17196 Mouse
- Entrez Gene: 24547 Rat
- Omim: 159430 Human
- SwissProt: P02687 Cow
- SwissProt: P02686 Human
- SwissProt: P04370 Mouse
see all -
Alternative names
- GDB antibody
- Golli MBP antibody
- Golli MBP; myelin basic protein antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] (ab209328)
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.
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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).
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ab209328 staining Myelin Basic Protein in SHSY5Y cells. The cells were fixed with 100% methanol (5 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 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195884, Rat monoclonal to alpha Tubulin (Alexa Fluor® 647), at a 1/250 dilution (shown in red).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] (ab209328)
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myelin Basic Protein antibody [IGX3421] (ab209328)
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.
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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.
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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.
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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 minutesThis 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab209328 has been referenced in 8 publications.
- Pandit K et al. An open source toolkit for repurposing Illumina sequencing systems as versatile fluidics and imaging platforms. Sci Rep 12:5081 (2022). PubMed: 35332182
- Ma L et al. Electroacupuncture-Regulated miR-34a-3p/PDCD6 Axis Promotes Post-Spinal Cord Injury Recovery in Both In Vitro and In Vivo Settings. J Immunol Res 2022:9329494 (2022). PubMed: 36132985
- Pasquettaz R et al. Peculiar protrusions along tanycyte processes face diverse neural and nonneural cell types in the hypothalamic parenchyma. J Comp Neurol 529:553-575 (2021). PubMed: 32515035
- Awad H et al. Endovascular repair and open repair surgery of thoraco-abdominal aortic aneurysms cause drastically different types of spinal cord injury. Sci Rep 11:7834 (2021). PubMed: 33837260
- Ravera S et al. Efficient extra-mitochondrial aerobic ATP synthesis in neuronal membrane systems. J Neurosci Res 99:2250-2260 (2021). PubMed: 34085315
- Tian C et al. Transient receptor potential ankyrin 1 contributes to the lysophosphatidylcholine-induced oxidative stress and cytotoxicity in OLN-93 oligodendrocyte. Cell Stress Chaperones 25:955-968 (2020). PubMed: 32572784
- Chen Z et al. Hyperglycemia aggravates spinal cord injury through endoplasmic reticulum stress mediated neuronal apoptosis, gliosis and activation. Biomed Pharmacother 112:108672 (2019). PubMed: 30784940
- Witt MR et al. Calcium homeostasis in fibroblasts from patients with amyotrophic lateral sclerosis. J Neurol Sci 126:206-12 (1994). PubMed: 7853028