Figure 1. GluN1 expression in cultured cortical neurons visualized using HB7535.
Figure 2. GluN1 histoblot in a horizontal rat brain section using HB7535.
Figure 3. GluN1 and NeuN expression in rat cerebellum mapped using HB7535.
Figure 4. GluN1 expression in brain P2 membrane fractions when probed with HB7535.
Figure 5. GluN1 expression in rat dentate gyrus mapped using HB7535.
Figure 6. Colocalization of GluN1 and GluA1-4 in cultured cortical rat neurons using HB7535.
Figure 7. GluN1 expression only observed within brain P2 membrane fractions when probed with HB7535.
Figure 8. GluN1 receptors on the dendrite of a cultured rat cortical neuron visualized using HB7535.
Figure 9. GluN1 histoblot in a horizontal rat brain section using HB7535.
Figure 10. Concentration response of HB7535 staining in a rat brain P2 preparation.
Figure 11. GluN1 histoblot in a horizontal rat brain section using HB7535.
Figure 1. GluN1 expression in cultured cortical neurons visualized using HB7535.
Figure 2. GluN1 histoblot in a horizontal rat brain section using HB7535.
Figure 3. GluN1 and NeuN expression in rat cerebellum mapped using HB7535.
Figure 4. GluN1 expression in brain P2 membrane fractions when probed with HB7535.
Figure 5. GluN1 expression in rat dentate gyrus mapped using HB7535.
Figure 6. Colocalization of GluN1 and GluA1-4 in cultured cortical rat neurons using HB7535.
Figure 7. GluN1 expression only observed within brain P2 membrane fractions when probed with HB7535.
Figure 8. GluN1 receptors on the dendrite of a cultured rat cortical neuron visualized using HB7535.
Figure 9. GluN1 histoblot in a horizontal rat brain section using HB7535.
Figure 10. Concentration response of HB7535 staining in a rat brain P2 preparation.
Figure 11. GluN1 histoblot in a horizontal rat brain section using HB7535.
Product information
Immunogen
Amino acids 1-564 of rat GluN1 expressed in a fusion protein
Epitope
Amino acids 341-561
Clone number
R1JHL
Isotype
IgG
Purification
Culture supernatent
Concentration
0.3mg/ml
Formulation
Lyophilised. When reconstituted contains PBS with 0.05% sodium azide and 1% recombinant albumin
Predicted species reactivity
Mouse, Rat, Human
Tested species reactivity
Mouse, Rat
Tested applications
Applications
WB, IHC(IF), Histoblot
Western blot optimal concentration
300ng/ml (1:1000 dilution) as tested in a rat brain P2 membrane preparation
IHC(IF) optimal concentration
300ng/ml (1:1000 dilution) as tested in rat brain hippocampal sections. Please note that utilisation of a citrate antigen retrieval protocol is required for successful staining.
ICC optimal concentration
300ng/ml (1:1000 dilution) as tested in cultured rat cortical neurons. Please note that utilisation of antigen retrieval is required for successful staining (10 minutes at 95°C in 100mM Tris, 5% urea, pH9.5).
Histoblot optimal concentration
300ng/ml (1:1000 dilution) as tested in horizontal rat brain sections
Product specific protocols
For IHC(IF) this antibody requires citrate antigen retrieval. For retrieval, incubate sections with 10mM citric acid, 0.05% Tween 20, pH6 at 95°C for 30 minutes followed by a 20 minute cooling period.
Positive control
GluN1 is widely expressed in the brain therefore neural tissues serve as an excellent positive control.
Negative control
Tissues such as the liver, heart and lung lack GluN1 expression while popular cell lines such as HeLa and HEK293 also lack expression therefore are good negative controls.
GluN1 has seven isoforms produced by alternative splicing:
Isoform 3 (canonical), known as Long isoform or NR1-3 - 938aa, 105.3kDa
Isoform 1, known as Short isoform or NR1-1 – 885aa, 99.3kDa
Isoform 2, known as Medium isoform or NR1-2 – 901aa, 101.2kDa
Isoform 4 – 922aa, 103.5kDa
Isoform 5 – 959aa, 107.9kDa
Isoform 6 – 943aa, 106.0kDa
Isoform 7 – 906aa, 101.9kDa
Expression
GluN1 is expressed alongside GluN2 as a heterotetramer in N-methyl-D-aspartate (NMDAR) receptors with each subunit containing two GluN1 subunits and a combination of two GluN2 (GluN2A, GluN2B, GluN2C or GluN2D) subunits. NMDAR receptors are expressed widely throughout the CNS and PNS. GluN1 expression has also been found in the kidney, heart and within bone alongside being reported in adipose tissue and the bladder.
Subcellular expression
GluN1 is expressed as part of NMDA receptors primarily within the post-synaptic densities found within the dendrites of neurones.
Processing
Following translation, the signal peptide (amino acids 1-18) is removed to leave the main peptide sequence.
Post translational modifications
GluN1 has multiple glycosylation sites alongside phosphorylation sites on residues 889, 890, 896 and 897.
Homology (compared to human)
Mouse GluN1 has a 99.04% homology to the human protein (9 amino acid changes) while Rat GluN1 has a 99.25% homology to human GluN1 (7 amino acid changes). Mouse and rat proteins have a 99.8% homology with only a single amino acid change (V460I).
Similar proteins
No other proteins with a significant homology were identified in a BLAST search. Other NMDAR subunits have <29% identity to GluN1.
Epitope homology (between species)
There is a 100% match between the epitope of HB7535 and human GluN1 while the mouse and rat proteins show 99.09% and 99.55% identity respectively.
Epitope homology (other proteins)
The only homology identified was with GluN3A and GluN3B however these were both of low similarity with identity scores of 33.1% and 33.2% respectively.
Storage & Handling
Storage instructions
-20°C then use reconstitution advice
Reconstitution advice
We recommend reconstituting with 50µl of either:
dH2O and storing at 4°C
50:50 ratio of dH2O to glycerol and storing at -20°C
dH2O then aliquot and store at -80°C
Take care when opening as the precipitate is extremely light and can easily be lost if disturbed. When reconstituting make sure that the antibody is thoroughly dissolved by pipetting up and down before giving the antibody a brief spin at <10,000g to make sure that all material is recovered and at the bottom of the tube.
Does this GluN1 antibody cross-react with other NMDA receptor subunits?
This clone has been reported to have no cross-reactivity with any GluN2 family subunits.
What guarantee do you have that my GluN1 antibody will perform as expected?
We guarantee that your GluN1 antibody will work for the applications and species we list on the datasheet. If the antibody fails to perform as expected then we are happy to offer a 100% refund guarantee. For more details please see our guarantee policy.
What protocols are available for use with this GluN1 antibody?
We have made a comprehensive collection of protocols that we have used in our experiments to validate this GluN1 antibody.
Will my GluN1 antibody work against species that have not been listed on the datasheet?
A species not being listed doesn’t mean that the GluN1 antibody won’t work, just that we haven’t tested it. If you test one of our antibodies in a new species please let us know (positive or negative)!
What counterstains do you recommend for use in ICC and IHC with this GluN1 antibody?
Western Blot Protocol (1 MB) 
