Murine Neuroinflammation Immunostaining Protocol

We obtained murine recombinant IL-33 (580506), AF647-conjugated Aβ (clone: 6E10) antibody (803021), APC-conjugated MHC-II (clone: M5/114.15.2) antibody (107614), FITC-conjugated VCAM1 (clone: MVCAM.A) antibody (105706) and MHC-II (I-A/I-E) (clone: M5/114.15.2) antibody (107601) from BioLegend. We obtained ICAM1-neutralizing (clone: YN1/1.7.4) antibody (BE0020)^{75 (link),76 (link)} and VCAM1-neutralizing (clone: M/K-2.7) antibody (BE0027)^{69 (link),77 (link)} from Bio X Cell. We obtained AF488-conjugated CD11b (clone: M1/70) antibody (53-0112-82), APC-conjugated CD11b (clone: M1/70) antibody (17-0112-83) and biotinylated CD11b (clone: M1/70) antibody (13-0112-82) from eBioscience. ApoE-neutralizing (clone: HJ6.3) antibody was a gift from D. Holtzman^{36 (link)}. We obtained DAPI (D3571) from Life Technologies, and mouse ITGB2 recombinant protein (LS-G14036-10) was from LSBio. We obtained mouse ApoE recombinant protein (MBS955382) from MyBioSource as well as CCR7 neutralizing (clone: 4B12) antibody (MAB3477)^{78 (link)} and VCAM1 antibody (BBA5) from R&D Systems. We obtained recombinant mouse CD44 protein (53953-M08H) from Sino Biological, MeX04 (4920) from Tocris Bioscience and Iba1 antibody (019-19741) from Wako.

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Lau S.F., Wu W., Wong H.Y., Ouyang L., Qiao Y., Xu J., Lau J.H., Wong C., Jiang Y., Holtzman D.M., Fu A.K, & Ip N.Y. (2023). The VCAM1–ApoE pathway directs microglial chemotaxis and alleviates Alzheimer’s disease pathology. Nature Aging, 3(10), 1219-1236.

Publication 2023

MHC-II (I-A/I-E) DAPI (D3571) Iba1 antibody

Af647 Antibody Apoe Biological Cd11b Cell Clone Dapi Fitc Icam1 Il 33 Mouse Mouse apoe protein Mouse cd44 protein Recombinant protein Sino

Corresponding Organization :

Other organizations : University of Hong Kong, Hong Kong University of Science and Technology, Hope Center for Neurological Disorders, Hong Kong Science and Technology Parks Corporation

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Variable analysis

independent variables

Murine recombinant IL-33 (580506)
ICAM1-neutralizing (clone: YN1/1.7.4) antibody (BE0020)
VCAM1-neutralizing (clone: M/K-2.7) antibody (BE0027)
ApoE-neutralizing (clone: HJ6.3) antibody
CCR7 neutralizing (clone: 4B12) antibody (MAB3477)

dependent variables

AF647-conjugated Aβ (clone: 6E10) antibody (803021)
APC-conjugated MHC-II (clone: M5/114.15.2) antibody (107614)
FITC-conjugated VCAM1 (clone: MVCAM.A) antibody (105706)
MHC-II (I-A/I-E) (clone: M5/114.15.2) antibody (107601)
AF488-conjugated CD11b (clone: M1/70) antibody (53-0112-82)
APC-conjugated CD11b (clone: M1/70) antibody (17-0112-83)
Biotinylated CD11b (clone: M1/70) antibody (13-0112-82)

control variables

DAPI (D3571)
Mouse ITGB2 recombinant protein (LS-G14036-10)
Mouse ApoE recombinant protein (MBS955382)
Recombinant mouse CD44 protein (53953-M08H)
MeX04 (4920)
Iba1 antibody (019-19741)

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