Log In Sign up
The largest database of trusted experimental protocols

Mab377

Manufactured by Abcam
Visit Supplier
Sourced in United States, Japan

MAB377 is a monoclonal antibody that recognizes the GFAP (Glial Fibrillary Acidic Protein) protein. GFAP is a type III intermediate filament protein that is expressed in astrocytes and other glial cells in the central nervous system.

Automatically generated - may contain errors

Lab products found in correlation

Ab5076, by Abcam (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Lsm 510, by Zeiss (4 mentions) Lsm 510 meta, by Zeiss (4 mentions) Ab7260, by Abcam (3 mentions) Odyssey infrared imaging system, by LI COR (3 mentions) Neuronal nuclei (neun), by Merck Group (3 mentions) Mab3402, by Merck Group (3 mentions) Target retrieval solution, by Agilent Technologies (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Ab125212, by Abcam (2 mentions) Ab4193, by Abcam (2 mentions) Diaminobenzidine, by Agilent Technologies (2 mentions) Z0334, by Agilent Technologies (2 mentions) Ba 2000, by Vector Laboratories (2 mentions) Avidin biotin peroxidase system, by Vector Laboratories (2 mentions) Stp 120 spin tissue processor, by Thermo Fisher Scientific (2 mentions) Rabbit anti ki67, by Cell Marque (2 mentions) Ba 1000, by Vector Laboratories (2 mentions) Ab53169, by Santa Cruz Biotechnology (2 mentions)

57 protocols using mab377

1

Immunostaining Protocol for Neuroinflammation Assessment

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunostaining was conducted following established protocols [23 (link)]. Briefly, brain tissues were collected after cardiac perfusion with cold 4% paraformaldehyde (PFA), placed in 4% PFA overnight, and then dehydrated using 30% sucrose/PBS for 4 days. Tissues at distances of − 2.5 to − 5 mm from bregma were selected for serial sections of 25-μm thickness. Sections were incubated overnight with primary antibody, including anti-NeuN (1:200, MAB377, Abcam), anti-cleaved caspase-3 (1:400, #9661, CST), anti-Iba-1 (1:1000, 019–19741, Wako, Japan), anti-Iba-1 (1:500, ab5076, Abcam), anti-CD68 (1:1000, ab125212, Abcam), and anti-CD206 (1:500, AF2535, R&D, USA), and then incubated for 2 h at room temperature with the appropriate fluorescent secondary antibody (Jackson ImmunoResearch Laboratories). DAPI (Southern Biotech) was utilized for nuclear staining and mounting. Micrographs were taken using a confocal microscope (Olympus, Japan). Three sections per rat were used, and three random fields in the ipsilateral basal cortex were acquired in each section, and the immunopositive cells in the basal cortex were quantified using Image J software [24 (link)].
Wang J., Wang L., Wu Q., Cai Y., Cui C., Yang M., Sun B., Mao L, & Wang Y. (2023). Interleukin-4 Modulates Neuroinflammation by Inducing Phenotypic Transformation of Microglia Following Subarachnoid Hemorrhage. Inflammation, 47(1), 390-403.
+ Open protocol
+ Expand
2

Immunofluorescence Staining of Brain Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols
Brain sections were washed with PBS for three times and were blocked with PBS-containing 3% goat serum (Vector Laboratories, #) and 0.1% Triton X-100 for 1 h at room temperature. Sections were incubated with primary antibodies overnight at 4°C and were washed with PBS. The following primary antibodies were used: GFAP (), mouse anti-Neuronal Nuclei (NeuN, Millipore, MAB377), DCX (), and GluA1 (Abcam, ab1232). On the second day, the samples were taken out and washed with PBS for 3 times, 5 min/time, followed by incubation with the secondary antibodies for 1 h at room temperature. Fluorophore-conjugated secondary antibody was used: goat anti-mouse Alexa Fluor 568 (Invitrogen, A11031), goat anti-rat Alexa Fluor 568 (Invitrogen, A11077), goat anti-rabbit Alexa Fluor 488 (Invitrogen, A11008), and goat anti-mouse Alexa Fluor 488 (Invitrogen, A11001). All the sections were observed and images were taken with a confocal microscope (Leica). The images were analyzed with Imarus software.
Xu W., Zhang X., Liang F., Cao Y., Li Z., Qu W., Zhang J., Bi Y., Sun C., Zhang J., Sun B., Shu Q, & Li X. (2021). Tet1 Regulates Astrocyte Development and Cognition of Mice Through Modulating GluA1. Frontiers in Cell and Developmental Biology, 9, 644375.
+ Open protocol
+ Expand
3

Immunophenotyping of LHX2+/NeuN- Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary antibodies used for LHX2+/NeuN−-based sorts included: Anti-LIM Homeobox 2 LHX2 (EMD Millipore, AB5756, 1:500), anti-NeuN (Millipore Sigma, MAB377, 1:2500), rabbit IgG isotype control (Abcam, ab172730, 1:1000), mouse IgG1 isotype control (Millipore Sigma, MABC002, 1:1250). Primary antibodies were detected with goat anti-rabbit IgG (H&L) secondary antibody conjugated to Alexa Fluor 488 (Invitrogen, A11034, 1:4,000) or goat anti-mouse IgG (H&L) secondary antibody conjugated to Alexa Fluor 647 (Invitrogen, A21235, 1:7,000), respectively. Gates were set to collect DAPI+, LHX2+, and NeuN− singlet events, and on average, ~88,000 DAPI+/LHX2+/NeuN− nuclei were captured per donor (see Table S3 for all LHX2+/NeuN−sorting outputs).
Sadick J.S., O’Dea M.R., Hasel P., Dykstra T., Faustin A, & Liddelow S.A. (2022). Astrocytes and oligodendrocytes undergo subtype-specific transcriptional changes in Alzheimer’s disease. Neuron, 110(11), 1788-1805.e10.
+ Open protocol
+ Expand
4

Immunohistochemical Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary antibodies used were: rabbit anti-DIP2B (1:2,000; Sigma HPA046133), mouse anti-α tubulin (1:5,000; Abcam ab7291), mouse anti-ace α tubulin (1:5,000; Sigma T7451), chicken anti-LacZ (1:2,000; Aves AB_2313508), mouse anti-tau1 (1:1,000; Abcam ab75714), rabbit anti-CamKII (1:500; Abcam ab52476), mouse anti-GABA (1:500; Sigma A0310), mouse anti-NeuN (1:800; Millipore MAB377), rabbit anti-GFAP (1:500; Abcam ab7260), chicken anti-GFP (1;2,000; Aves AB_2307313), mouse anti-GAPDH (1:5,000; Transgen HC301-01), rabbit anti-GST (1:2,000; CST 2625), mouse anti-His (1:2,000; Abcam ab18184).
Secondary antibodies used were: anti-Mouse IgG-HRP (1:5,000; Santa Cruz Biotechnology sc-2005), Goat-anti-Rabbit IgG-HRP (1:5,000; Invitrogen PI31460), Donkey anti-Mouse Alexa Fluor 488 (1:5,000; Invitrogen A21202), Goat anti-Chicken Alexa Fluor 488 (1:5,000, Invitrogen A11039), Donkey anti-Rabbit Alexa Fluor 546 (1:5,000, Invitrogen A10040), Donkey anti-Mouse Alexa Fluor 546 (1:5,000, Invitrogen A10036).
Xing Z.K., Zhang L.Q., Zhang Y., Sun X., Sun X.L., Yu H.L., Zheng Y.W., He Z.X, & Zhu X.J. (2020). DIP2B Interacts With α-Tubulin to Regulate Axon Outgrowth. Frontiers in Cellular Neuroscience, 14, 29.
+ Open protocol
+ Expand
5

Histological and Immunohistochemical Analysis of Mouse Brain

Check if the same lab product or an alternative is used in the 5 most similar protocols
For histological and immunohistochemical analyses, brains from treated mice were fixed in 4% phosphate-buffered formaldehyde, dehydrated in a STP 120 Spin Tissue Processor (Thermo Scientific) and embedded in paraffin. Sections (5μm thickness) were cut and mounted onto glass slides, deparaffinized in xylene and rehydrated in a descending series of alcohols, then rinsed in distilled water and stained. Hematoxylin-eosin (HE) staining was carried out according to routine laboratory protocols. For immunohistochemical stainings, enzyme-induced antigen retrieval (for anti-GFAP) using pepsin (1% in 0,01 M HCl) or heat-induced antigen retrieval (all other antibodies) using a 1x target retrieval solution (Dako) was performed. Primary antibodies used were rabbit anti-GFAP (Dako Z0334, 1:1000), rabbit anti-Ki67 (CellMarque, Rocklin, CA, 275R-14, 1:200), mouse anti-NeuN (Merck Millipore, Billerica, Ma, MAB377, 1:200), and rabbit anti-SFRP1 (Abcam, ab4193, 1:200). Secondary antibodies were diluted 1:500 (anti-mouse, VectorLab, BA-2000 or anti-rabbit, VectorLab, BA-1000) and staining was visualized using the avidin-biotin peroxidase system (VectorLab) and freshly prepared diaminobenzidine as chromogen (Dako). Slides were counterstained with haematoxylin, dehydrated and mounted.
Zuckermann M., Hovestadt V., Knobbe-Thomsen C.B., Zapatka M., Northcott P.A., Schramm K., Belic J., Jones D.T., Tschida B., Moriarity B., Largaespada D., Roussel M.F., Korshunov A., Reifenberger G., Pfister S.M., Lichter P., Kawauchi D, & Gronych J. (2015). Somatic CRISPR/Cas9-mediated tumour suppressor disruption enables versatile brain tumour modelling. Nature Communications, 6, 7391.
+ Open protocol
+ Expand
6

Comprehensive Immunostaining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunofluorescence staining analysis was performed on 40 μm free-floating sections. Primary antibodies, including chicken anti-GFP (Aves Labs, GFP-1020), goat anti-CD206 (R&D Systems, AF2535), rabbit anti-Cleaved Caspase-3 (Cell Signaling, 9661), mouse anti-Caspase-3 (Novus Biologicals, 31A1067), mouse anti-NeuN (Millipore, MAB377), rabbit anti-Tmem119 (Abcam, ab209064), mouse anti-LPL (Abcam, ab21356), rabbit anti-CX3CR1(Abcam, ab8021), rabbit anti-GFAP (Abcam, ab7260), mouse anti-iNOS (inducible nitric oxide synthase) (Abcam, ab49999), and goat anti-Iba1 (Abcam, ab5076), were used. Corresponding secondary antibodies, including 488- and 594-conjugated secondary antibodies, were purchased from Thermo Fisher Scientific, Alexa Fluor conjugates.
All images were processed with Image J for quantification analysis. The means were calculated from 3 randomly selected microscopic fields in the ipsilateral and contralateral cortex of each section, respectively, and 3 consecutive sections were analyzed for each brain. Data are expressed as mean numbers of cells per square millimeter.
Ye S.Y., Apple J.E., Ren X., Tang F.L., Yao L.L., Wang Y.G., Mei L., Zhou Y.G, & Xiong W.C. (2019). Microglial VPS35 deficiency regulates microglial polarization and decreases ischemic stroke-induced damage in the cortex. Journal of Neuroinflammation, 16, 235.
+ Open protocol
+ Expand
7

Immunofluorescence Staining of Neural Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were washed with PBS, fixed with 4% (w/v) paraformaldehyde, blocked with 5% (v/v) BSA in PBS for 30 min, and permeabilized with 0.1% (v/v) Triton X-100 in PBS for 5 min or digitonin for 30 min. Primary antibody to NGN2 (Abnova H00063973-M10), NeuN (Millipore MAB377), MAP2 (abcam ab32454) and αSyn (clone 15G7, Enzo Life Sciences ALX-804-258-LC05) was incubated for 1 h and washed with PBS; the secondary antibody conjugated to Alexa Fluor dye was incubated for 1 h, washed with PBS, and visualized by confocal microscopy (Zeiss LSM710).
Sanyal A., Novis H.S., Gasser E., Lin S, & LaVoie M.J. (2020). LRRK2 Kinase Inhibition Rescues Deficits in Lysosome Function Due to Heterozygous GBA1 Expression in Human iPSC-Derived Neurons. Frontiers in Neuroscience, 14, 442.
+ Open protocol
+ Expand
8

Lentiviral Delivery of Dominant-Negative IκBα

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Lentiviral IκBα was cloned by inserting cDNA of dominant-negative IκBα in synapsin promoter-driven lentiviral vector as previously described14 (link), and replacement of IκBα by GFP cDNA was used as the matched control. Lentiviruses were generated in HEK293T cells and then purified as previously described14 (link). Brain histology was analyzed using brain sections and immunostaining. Mice under anesthesia were transcardially perfused with 4% PFA and brains were removed, post-fixed in 4% PFA for four hours, and infiltrated with 20% – 30% sucrose. 20 μm-thick brain sections were blocked with serum of appropriate species, penetrated with 0.2% Triton-X 100, treated with primary antibodies including mouse anti-GFAP (Millipore, MAB3402, 1:1000), mouse anti-NeuN (Millipore, MAB377, 1:1000), rabbit anti-TGF-β (Abcam, ab53169, 1:200), and mouse anti-HuR (Santa Cruz, sc5261, 1:500), and subsequently reacted with fluorescent secondary antibodies (Invitrogen, 1:1000). Naïve IgGs of appropriate species were used as negative controls. DAPI staining was used to reveal all cells in the section. Images were taken using a confocal microscope.
Yan J., Zhang H., Yin Y., Li J., Tang Y., Purkayastha S., Li L, & Cai D. (2014). Obesity- and aging-induced excess of central transforming growth factor-β potentiates diabetic development via an RNA stress response. Nature medicine, 20(9), 1001-1008.
+ Open protocol
+ Expand
9

Immunocytochemistry and Immunostaining Protocol

Cited 3 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunocytochemistry and immunostaining were conducted according to published approaches (Guo et al., 2011 (link), Liu et al., 2010 (link), Liu et al., 2013a (link)). For immunostaining cultured cells, anti-neuron-specific type β-III tubulin (Biolegend, #801202; 1:1,000), anti-glial fibrillary acidic protein (Millipore, MAB377; 1:1,000), anti-BrdU (Abcam, ab6326; 1:2000), and anti-cleaved caspase 3 (Cell Signaling, #9664; 1:200) were used as the primary antibodies. For immunohistochemistry staining, the primary antibodies used were as follows: anti-Ki67 (Thermo Fisher, RM9106; 1:1,000), anti-BrdU (Abcam, ab6326; 1:1,000), and anti-DCX (Cell Signaling, #4604; 1:500). The secondary antibodies conjugated to Alexa Fluor 488 or 594 with a concentration of 1:500 were used at room temperature. To analyze the amount of BrdU+GFP+, or BrdU+DCX+GFP+ cells in the DG, we used one in six series of 40-μm brain sections starting at beginning of hippocampus (relative to bregma, −1.5 mm) to the end of hippocampus (relative to bregma, −3.5 mm). For details, see Supplemental Experimental Procedures.
Liu P.P., Tang G.B., Xu Y.J., Zeng Y.Q., Zhang S.F., Du H.Z., Teng Z.Q, & Liu C.M. (2017). MiR-203 Interplays with Polycomb Repressive Complexes to Regulate the Proliferation of Neural Stem/Progenitor Cells. Stem Cell Reports, 9(1), 190-202.
+ Open protocol
+ Expand
10

Immunofluorescence Staining of Nervous System

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The following antibodies were used: GFP (1:500, Life Technologies, A10262 or A11122); Nefh (1:1,000, Aves laboratory, NFH or 1:1,000, Abcam, ab8135); CGRP (1:500, Abcam, ab43873); NeuN (1:500, Millipore, MAB377); PECAM1 (1:100, Abcam, ab28364); PV (1:500, Swant, PV 2); α-bungarotoxin (1:500, Life Technologies, B13423); advillin (1:500, Abcam, ab72210); Tuj1 (1:500, Covance, MMS-435P-250); αSMA–Cy3 (1:200, Sigma, C6198); E-cadherin (1:500, Life Technologies, 13–1900); MUC-1 (1:500, Fisher Scientific, HM1630P0); T1α (1:50, DSHB, 8.1.1); proSP-C (1:300, Millipore, AB3786); ChAT (1:300, Millipore, AB143); Piezo2 (1:1,000)21 (link); IB4-Alexa Fluor568 conjugate (1:200, Life Technologies, I21412).
Nonomura K., Woo S.H., Chang R.B., Gillich A., Qiu Z., Francisco A.G., Ranade S.S., Liberles S.D, & Patapoutian A. (2016). Piezo2 senses airway stretch and mediates lung inflation-induced apnoea. Nature, 541(7636), 176-181.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!