PA5-28128

antibody from Invitrogen Antibodies

Targeting: ADAM33 C20orf153, dJ964F7.1, DKFZp434K0521

Provider product page for PA5-28128

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Other assay [2]

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Product number: PA5-28128 - Provider product page
Provider: Invitrogen Antibodies
Product name: ADAM33 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Recommended positive controls: A431, H1299, HeLaS3, Raji. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.7 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ADAM33 protein structure - PA5-28128 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Extended Data Figure 3 | Immunohistochemical validations of differentially expressed genes in the thalamus of Grn -/- mice. a-b. Validations using immunohistochemistry and confocal microscopy confirm the downregulation of P2Y12 and Tmem119 in Grn -/- thalamic microglia at 12 and 19 months, respectively (panel a). In contrast, Grn -/- thalamic microglia show marked increases in ApoE and Adam33 protein detected by immunohistochemical staining and confocal microscopy (panel b). Insets are high magnification images from the boxed areas in the ventral thalamus. Confocal images on the right panels are obtained from 12 months old Grn +/+ and Grn -/- thalamus. Immunohistochemistry was performed in 3 independent mice per genotype, whereas the confocal images were from two independent mice. c. Confocal images showing upregulated expression of Cathepsin B, IGF-1 and GPNMB in 12 months old Grn -/- thalamic microglia. In contrast, Grn -/- thalamic microglia show reduced expression of Numb. The validations were performed in N=3 independent mice per genotype with similar results. d. A proposed model showing the age-dependent transition of Grn -/- thalamic microglia from a homeostatic state to disease state from 7 to 19 months. The defects in Grn -/- microglia downregulate homeostatic genes ( C1qa , C1qb , Mef2c , Csf1r , Cx3cr1 , Tgfbr1 , Tmem119 , Adam33 , Igf1 , P2ry12 ), and upregulate genes related to lysosomal functions ( Ctsb ), lipid transport ( Apoe ), intracellular trafficking ( My

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Extended Data Figure 5 | Characterization of P3 primary microglia from Grn +/+ and Grn -/- mice using single cell RNA-seq, NanoString nCounter neuroinflammation panel, and western blots. a. A schematic diagram illustrating the study design to characterize primary microglia from postnatal day 3 (P3) Grn +/+ and Grn -/- mice using scRNA-seq and NanoString nCounter neuroinflammation panel, and to prepare serum-free conditioned media from Grn +/+ and Grn -/- P3-MG. In parallel, primary cortical neurons and GABAergic inhibitory neurons are isolated from the developing cortex and ganglionic eminences of embryonic day 15.5 (E15.5) Grn +/+ and Grn -/- mice. After 14 days in vitro (DIV), Grn +/+ and Grn -/- microglial conditioned media (MCM) are added to Grn +/+ and Grn -/- excitatory neurons or GABAergic inhibitory neurons and incubate for 24 hours. b. t -SNE plots of scRNA-seq data from Grn +/+ and Grn -/- P3-MG revealed 4 distinct clusters and the extent of overlapping in cell density and cluster distribution between Grn +/+ and Grn -/- P3-MG. c. Comparison of clusters A of P3-MG with 2 to 19 months (mo) thalamic microglia (Th-MG) reveals more overlapping between P3-MG (black) and 19mo Th-MG (red). d. Hierarchical clustering of gene expression in Grn +/+ and Grn -/- P3-MG cluster A and 19 months old Th-MG. e. Venn diagrams showing the extent of overlapping between DEGs from 12 and 19 months old Th-MG and DEGs in P3-MG identified by scRNA-seq (upper panel) or DEGs in P3-MG identifie