Yes, it is possible to use two primary antibodies from different species but of the same isotype for double staining Immunofluorescence. Specifically, for MAB397 and GluR2, one can use an anti-mouse and an anti-rabbit secondary antibody to distinguish reactivity of the primary antibodies.
MAB397
Anti-Glutamate Receptor 2 Antibody, extracellular, clone 6C4
clone 6C4, Chemicon®, from mouse
Synonym(s):
GluR2
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100 μG
€ 693,00
€ 693,00
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About This Item
UNSPSC Code:
12352203
NACRES:
NA.41
eCl@ss:
32160702
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Product Name
Anti-Glutamate Receptor 2 Antibody, extracellular, clone 6C4, clone 6C4, Chemicon®, from mouse
biological source
mouse
conjugate
unconjugated
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
6C4, monoclonal
species reactivity
monkey
species reactivity (predicted by homology)
mouse, rat, canine
manufacturer/tradename
Chemicon®
technique(s)
ELISA: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
radioimmunoassay: suitable
western blot: suitable
isotype
IgG2a
NCBI accession no.
UniProt accession no.
target post-translational modification
unmodified
Quality Level
Gene Information
dog ... Gria2(482667)
mouse ... Gria2(14800)
rat ... Gria2(29627)
rhesus monkey ... Gria2(574344)
1 of 4
This Item | MAB397A4 | MABN71 | MABN832 |
|---|---|---|---|
| clone 6C4, monoclonal | clone 6C4, monoclonal | clone L21/32, monoclonal | clone 2D8, monoclonal |
| species reactivity monkey | species reactivity mouse, rat | species reactivity rat | species reactivity human, rat |
| biological source mouse | biological source mouse | biological source mouse | biological source mouse |
| antibody form purified immunoglobulin | antibody form purified immunoglobulin | antibody form purified antibody | antibody form purified antibody |
| Gene Information dog ... Gria2(482667) | Gene Information human ... GRIA2(2891) | Gene Information human ... GRIA2(2891) | Gene Information human ... GRIA1(2890) |
| UniProt accession no. | UniProt accession no. | UniProt accession no. | UniProt accession no. |
Analysis Note
Control
Positive Control: Cerebral cortex and pyramidal neurons, mouse brain lysate.
Positive Control: Cerebral cortex and pyramidal neurons, mouse brain lysate.
Routinely evaluated by Western Blot on mouse brain lysates.
Western Blot Analysis:
1:1000 dilution of this lot detected glutamate receptor 2 on 10 μg of mouse brain lysates.
Western Blot Analysis:
1:1000 dilution of this lot detected glutamate receptor 2 on 10 μg of mouse brain lysates.
Application
Anti-Glutamate Receptor 2 Antibody, extracellular, clone 6C4 detects level of Glutamate Receptor 2 & has been published & validated for use in ELISA, IC, IH, IP, RIA & WB with more than 50 product citations.
Immunocytochemistry:
on 4% paraformaldehyde fixed cells was used in a previous lot:2-3 µg/mL (Vissavajjhala, 1996; Osten, 1998; Passafaro, 2001).
ELISA/RIA:
A previous lot of this antibody was used in ELISA/RIA.(Vissavajjhala, 1996).
Immunohistochemistry on 50 µm, 4% paraformaldehyde fixed brain sections: 1:500-1:800 (Vissavajjhala, 1996; Yung, 1998; Kumar; 2002).
Immunoprecipitation: 2-4 µg/mL (Osten, 1998).
Western blot: 1-2 µg/mL (Vissavajjhala, 1996; Osten, 1998); membrane preparations are suggested for enhanced signals.
Optimal working dilutions and protocols must be determined by end user.
on 4% paraformaldehyde fixed cells was used in a previous lot:2-3 µg/mL (Vissavajjhala, 1996; Osten, 1998; Passafaro, 2001).
ELISA/RIA:
A previous lot of this antibody was used in ELISA/RIA.(Vissavajjhala, 1996).
Immunohistochemistry on 50 µm, 4% paraformaldehyde fixed brain sections: 1:500-1:800 (Vissavajjhala, 1996; Yung, 1998; Kumar; 2002).
Immunoprecipitation: 2-4 µg/mL (Osten, 1998).
Western blot: 1-2 µg/mL (Vissavajjhala, 1996; Osten, 1998); membrane preparations are suggested for enhanced signals.
Optimal working dilutions and protocols must be determined by end user.
Biochem/physiol Actions
Recognizes the large N-terminal extracellular domain of Glutamate Receptor 2 (GluR2). No cross-reactivity observed with other AMPA/Kainate GluR subunits. On western blots of brain extracts from rat, macaque monkey, and dog, MAB397 recognizes a band at approximately 102 kDa corresponding to full length GluR2. Other proteins noted by Western blot at 66 kDa or lower molecular weight appear to be breakdown products of GluR2 (Vissavajjhala, 1996). By immunohistochemistry GluR2 is widely distributed at both the cellular and synaptic levels. MAB397 recognizes GluR2 present in a vast majority of, but not all, GABAergic interneurons (Vissavajjhala, 1996).
General description
Glutamate receptors (GluRs) can be categorized as ionotropic or metabotropic and subcatergorized by their agonist preferences (NMDA, AMPA or Kainic acid). There are four types of AMPA selective GluR subunits (GluR1, GluR2, GluR3 and GluR4). Tetrameric or pentameric combinations of different subunits contributes to the functional diversity of AMPA receptors. In general, AMPA receptors mediate fast synaptic current at most excitatory synapses, with stoichiometry characterized by subtype composition. Although subunit composition of AMPA receptors varies, they must contain at least one edited GluR2 subunit to be calcium impermeable. The critical residue controlling calcium permeability is in the pore loop region. In GluR1, GluR3, and GluR4, this positionis occupied by a Gln residue. In GluR2, it is occupied by an Arg residue. It has been shown experimentally that the presence of Arg in this position blocks CA2+ ion permeability, while a Gln does not. Relative calcium permeability in AMPA receptor channels may be significant in pathological neurotoxic damage and long term changes in nervous system responses.
102 kDa
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Physical form
Format: Purified
Purified immunoglobulin from culture supernatant
Purified mouse immunoglobin IgG2a liquid in buffer containing PBS, no preservative.
Preparation Note
Stable for 6 months at -20ºC in undiluted aliquots from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage the IgG2a and affect product performance.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage the IgG2a and affect product performance.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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Description
Pricing
Storage Class
12 - Non Combustible Liquids
wgk
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Find documentation for the products that you have recently purchased in the Document Library.
S-SCAM/MAGI-2 is an essential synaptic scaffolding molecule for the GluA2-containing maintenance pool of AMPA receptors.
Danielson, E; Zhang, N; Metallo, J; Kaleka, K; Shin, SM; Gerges, N; Lee, SH
The Journal of Neuroscience null
Tyrosine phosphatases regulate AMPA receptor trafficking during metabotropic glutamate receptor-mediated long-term depression.
Moult, PR; Gladding, CM; Sanderson, TM; Fitzjohn, SM; Bashir, ZI; Molnar, E; Collingridge, GL
The Journal of Neuroscience null
Differential regulation of AMPA receptor and GABA receptor trafficking by tumor necrosis factor-alpha.
Stellwagen, D; Beattie, EC; Seo, JY; Malenka, RC
The Journal of Neuroscience null
Reuben Saba et al.
Molecular and cellular biology, 32(3), 619-632 (2011-12-07)
The dynamic expression of AMPA-type glutamate receptors (AMPA-R) at synapses is a key determinant of synaptic plasticity, including neuroadaptations to drugs of abuse. Recently, microRNAs (miRNAs) have emerged as important posttranscriptional regulators of synaptic plasticity, but whether they target glutamate
Regeneration of the adult zebrafish brain from neurogenic radial glia-type progenitors.
Kroehne, V; Freudenreich, D; Hans, S; Kaslin, J; Brand, M
Development null
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Glutamate is an excitatory neurotransmitter found in the synaptic vesicles of glutamatergic synapses. The post-synaptic neurons in these synapses contain ionotropic and metabotropic glutamate receptors. Glutamate binds to AMPA (α-amino-3-hydroxy-5- methylisoxazole-4-propionic acid) subtype glutamate receptors, leading to sodium influx into the post-synaptic cell and resulting in neuronal excitability and synaptic transmission. The NMDA (N-methyl-d-aspartate) subtype glutamate receptors, on the other hand, regulate synaptic plasticity, and can influence learning and memory. The metabotropic g-protein coupled mGluRs modulate downstream calcium signaling pathways and indirectly influence the synapse’s excitability. The synaptic architecture includes intracellular scaffolding proteins (PSD-95, GRIP), intercellular cell adhesion molecules (NCAMs, N-Cadherins), and a variety of signaling proteins (CaMKII/PKA, PP1/PP2B). Processes critical for synaptic transmission and plasticity are influenced by these molecules and their interactions. When the function of these molecules is disrupted, it leads to synaptic dysfunction and degeneration, and can contribute to dementia as seen in Alzheimer’s disease.
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Is it possible to use two primary antibodies from different species but of the same isotype for double staining Immunofluorescence? Specifically, considering MAB397 and GluR2.
1 answer-
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