PA5-28128

antibody from Invitrogen Antibodies

Targeting: ADAM33 C20orf153, dJ964F7.1, DKFZp434K0521

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-28128

Provider

Invitrogen Antibodies

Product name

ADAM33 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: A431, H1299, HeLaS3, Raji.

Concentration

0.7 mg/mL

References

Submitted references

Neurotoxic microglia promote TDP-43 proteinopathy in progranulin deficiency.

Zhang J, Velmeshev D, Hashimoto K, Huang YH, Hofmann JW, Shi X, Chen J, Leidal AM, Dishart JG, Cahill MK, Kelley KW, Liddelow SA, Seeley WW, Miller BL, Walther TC, Farese RV Jr, Taylor JP, Ullian EM, Huang B, Debnath J, Wittmann T, Kriegstein AR, Huang EJ
Nature 2020 Dec;588(7838):459-465

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using ADAM33 Polyclonal Antibody (Product # PA5-28128). Sample (30 µg of whole cell lysate). A: Raji. 7.5% SDS PAGE. ADAM33 Polyclonal Antibody (Product # PA5-28128) diluted at 1:500.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

ADAM33 Polyclonal Antibody detects ADAM33 protein at cytoplasm on human placenta by immunohistochemical analysis. Sample: Paraffin-embedded placenta. ADAM33 Polyclonal Antibody (Product # PA5-28128) dilution: 1:100. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Other assay

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