DLG4 antibody

Catalog No. ABIN361694

This independently validated Mouse Monoclonal antibody specifically detects DLG4 in WB, IHC, ICC, IF, AA and mIHC. It exhibits reactivity toward Rat.

Quick Overview for DLG4 antibody (ABIN361694)

Target: DLG4 (Discs, Large Homolog 4 (Drosophila) (DLG4))

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This DLG4 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Antibody Array (AA), Multiplex Immunohistochemistry (mIHC)

Clone: 6G6

Product Details anti-DLG4 Antibody

Specificity: Detects ~100 kDa. An additional protein of >100 kDa is also detected. Additional cross-reactive bands are detected at ~75 kDa and 50 kDa in rat and mouse samples.

Cross-Reactivity: Cow, Human, Mouse, Rat

Purification: Protein G Purified

Immunogen: Recombinant rat PSD-95

Isotype: IgG2a

Application Details

Application Notes:

• WB (1:250)
• IHC (1:1000)
• ICC/IF (1:100)
• optimal dilutions for assays should be determined by the user.

Comment:

1 μg/ml was sufficient for detection of PSD-95 on 20 μg rat brain tissue extract by ECL immunoblot analysis using Goat Anti-Mouse IgG: HRP as the secondary.

Restrictions: For Research Use only

Independent Validation (mIHC)

Validation #104331 (Multiplex Immunohistochemistry)

by: Akoya Biosciences

No.: #104331

Date: 04/20/2021

Antigen: DLG4

Lot Number: MP686150

Method validated: Multiplex Immunohistochemistry

Positive Control:

Fresh frozen mouse olfactory bulb

Negative Control:

Unlabeled control (mouse fresh frozen)

Notes:

Passed. The anti-DLG4 antibody ABIN361694 specifically labels areas that contain high levels of post-synaptic boutons; these are located throughout the murine olfactory bulb and are concentrated in the glomerular layer.

Validation Images

Murine fresh frozen coronal olfactory bulb section (Thickness = 5 µm) labeled with anti-DLG4 antibody ABIN361694 (red; bound to fluorophore ATTO 550). Labeling is present throughout olfactory bulb layers with a strong degree of labeling in the glomerular layer; this is consistent with the large number of postsynaptic targets within the glomerular neuropil. Slc17a7 and Map2 were labeled with ABIN1027710 (green; bound to fluorophore ATTO 550) and ABIN125739 (blue; bound to fluorophore ATTO 550).

FFPE normal human cortex tissue section labeled with anti-DLG4 antibody ABIN361694 (green; bound to fluorophore ATTO 550) after EDTA antigen retrieval. MAP2 and Synapsin were labeled with anti-MAP2 antibody ABIN125739 (cyan; bound to fluorophore ATTO 550) and anti-SYN1 antibody ABIN5542390 (red; bound to fluorophore AF488).

Full Methods

Primary Antibody: ABIN361694

Full Protocol:

• Protocol details are described in the Akoya Biosciences CODEX® User Manual (see https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf).
• Tissue preparation as outlined in the Akoya Biosciences CODEX® User Manual fresh-frozen tissue protocol.
• Conjugation of the anti-DLG4 antibody ABIN361694 to an oligo barcode used to bind oligo-conjugated fluorophore ATTO 550.

Experimental Notes:

• No signal was detected in unlabeled specimens.

• Specific staining of DLG4 was also observed with human FFPE cortical tissue sections with both citrate antigen retrieval and EDTA antigen retrieval.

• Optimal staining and signal to noise ratios were obtained if tissue was pre-treated for enogenous tissue autofluorescence removal (see https://www.akoyabio.com/wp-content/uploads/2020/07/Customer-Demonstrated-Protocol-Autofluorescence-Quenching-Mar2020.pdf).

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS pH 7.4, 50 % glycerol, 0.09 % sodium azide, Storage buffer may change when conjugated

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: -20°C

Target Details for DLG4

Target: DLG4 (Discs, Large Homolog 4 (Drosophila) (DLG4))

Alternative Name: PSD95

Background: Postsynaptic Density protein 95 (PSD95), also known as Synapse associated protein 90 kDa, is a member of the membrane-associated guanylate kinase (MAGUK) family of proteins. PSD95 is a scaffolding protein and is involved in the assembly and function of the postsynaptic density complex (1). These family members consist of an N-terminal variable segment followed by three amino-terminal PDZ domains, an upstream SH3 domain and an inactive carboxyl-terminal guanylate kinase (GK) domain. The first and second PDZ domain localize NMDA receptors and K+ channels to synapses, and the third binds to neuroligins which are neuronal cell adhesion molecules that interact with b-neurexins and form intercellular junctions. PSD-95 also binds to neuronal nitric oxide synthase, possibly through interactions between PDZ domains present on both proteins (2). Thus different PDZ domains of PSD-95 might be specialized for distinct functions (3, 4). PSD95 participates in synaptic targeting of AMPA receptors through an indirect manner involving Stargazin and related transmembrane AMPA receptor regulatory proteins (TARPs) (5). The protein is implicated in experience dependent plasticity and plays an indispensable role in learning (6). Mutations in PSD95 are associated with autism (7).

Gene ID: 29495

NCBI Accession: NP_062567

UniProt: P31016

Pathways: Regulation of Muscle Cell Differentiation, Synaptic Membrane, Skeletal Muscle Fiber Development, Asymmetric Protein Localization, Regulation of long-term Neuronal Synaptic Plasticity