Anti-NEFH Antibody (Monoclonal, N52)

IHC analysis of NEFH using anti-NEFH antibody (MA1071).
NEFH was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-NEFH Antibody (MA1071) overnight at 4°C. Peroxidase Conjugated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

Western blot analysis of NEFH using anti-NEFH antibody (MA1071).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-NEFH antigen affinity purified monoclonal antibody (Catalog # MA1071)) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for NEFH at approximately 200KD. The expected band size for NEFH is at 112KD.

IHC analysis of NEFH using anti-NEFH antibody (MA1071).
NEFH was detected in paraffin-embedded section of human glioma tissues. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-NEFH Antibody (MA1071) overnight at 4°C. Peroxidase Conjugated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

Spared nerve injury-induced mechanical hypersensitivity and Nav1.7 activation in rat injured DRGs. ( A , B ) Ipsilateral ( A ) and contralateral ( B ) paw withdrawal threshold (PWL) before or after SNI or sham surgery. ** P < 0.01 vs. the baseline (BL). n = 6 rats/group. Two-way ANOVA followed by post hoc Tukey, F 1,5 = 22.63 in ( A ), F 1,5 = 1.809 in ( B ). ( C ) Levels of SCN9A mRNA in L4-6 DRGs at days shown after SNI or sham surgery in rats. n = 3 rats per group per time point. One-way ANOVA followed by post hoc Tukey, F 3,8 = 51.67 for SNI group, F 3,8 = 2.026 for sham group. ( D ) Levels of Nav1.7 protein in L4-6 DRGs at days shown after SNI surgery in rats. One-way ANOVA followed by post hoc Tukey, F 3,8 = 0.9785 in contralateral side (Cont), F 3,8 = 109.7 in ipsilateral side (Ipsi) * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the zero day. n = 3 for each time point. ( E ) Co-localization of Nav1.7 with CGRP, IB4, and NF200 in rat L5 DRG, respectively. Scale bar: 100 μm.
Index in PubMed under a CC BY license. PMID: 30425258

Immunofluorescence identification of SCNs. (A) Cell nuclei are identified with DAPI. (B) SCNs are stained with the anti-NF200 monoclonal antibody. More than 90% of the cultured cells show positive expression of NF200 on the day 7 of ex vivo culture. The number of NF200-positive neurons are counted per area and expressed as percentage of total number of cells (Scale bar: 40 μm).
Index in PubMed under a CC BY license. PMID: 33425964

LIPUS improves cerebral WM integrity on day 28 after stroke. (A) Schematic of the localization of the peri-infarct cortex (CTX), external capsule (EC), and striatum (STR). (B) Double-labeled immunofluorescence staining of MBP (green) and SMI32 (red) in 3 brain regions, CTX, EC, and STR, on day 28 after dMCAO (i). Quantification of MBP immunoreactivity (ii) and the SMI32/MBP fluorescence intensity ratio (iii) are presented as the ratio of fold change compared to the Sham control, scale bar = 50 μm, n = 6 mice/group. (C) Representative images depicting that the Western blot analysis of MBPs was obtained for each group (i). The expression levels of the target proteins were normalized relative to β-actin (ii). (D) Representative immunostaining of MBP and SMI32 on day 28 after dMCAO (i), scale bar = 200 μm; (ii) quantification of the ratio of MBP to SMI32 staining-positive areas in ipsilateral injured brain tissue. Data were normalized to stained areas in the contralateral hemisphere, n = 6 mice/group. (E) Representative images of myelin coverage (i), calculated MBP+ myelin coverage area along NF200+ nerve fibers (ii), scale bar = 20 μm, n = 6 mice/group. Data are presented as means ± SD. Statistical analysis for (B) was conducted using 2-way ANOVA and then Tukey’s multiple comparisons test, while (C) and (E) were analyzed using one-way ANOVA and then Tukey’s multiple comparisons test. Analysis for (D) involved unpaired 2-tailed Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001,**** P < 0.0001.
Index in PubMed under a CC BY license. PMID: 40290135