711025

antibody from Invitrogen Antibodies

Targeting: NEFH NF-H, NFH

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

711025

Provider

Invitrogen Antibodies

Product name

NF-H Recombinant Polyclonal Antibody (4HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Cat, Dog, Monkey and Bovine.

Antibody clone number

4HCLC

Concentration

0.5 mg/mL

References

Submitted references

Chitosan Tubes Inoculated with Dental Pulp Stem Cells and Stem Cell Factor Enhance Facial Nerve-Vascularized Regeneration in Rabbits.

Mu X, Liu H, Yang S, Li Y, Xiang L, Hu M, Wang X
ACS omega 2022 Jun 7;7(22):18509-18520

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell and tissue extracts (30 µg lysate) of Rat Brain (Lane 1), Mouse Brain (Lane 2), Neuro-2a (Lane 3), Hek-293 (Lane 4), Hela (Lane 5), K562 (Lane 6), SH-SY5Y (Lane 7) and SH-SY5Y treated with Retinoic acid (10 uM/ 48hours) (Lane 8). The blots were probed with Anti-Neurofilament-H Recombinant Rabbit Polyclonal Antibody (Product # 711025, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 180 and 220 kDa band corresponding to Neurofilament-H isoforms was observed across tissues and cell lines tested according to the given treatment. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, retinoic acid (10 uM, 3days) treated SH-SY5Y cells were fixed and permeabilized for detection of Neurofilament-H using Neurofilament-H Recombinant Rabbit Polyclonal Antibody (Product # 711025, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Neurofilament-H protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of Neurofilament-H. Panel e) shows untreated cells with nuclear localization. Panel f) represents control cells with no primary antibody to assess background.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of the neurofilament heavy chains in frozen sections of human cortex. Tissues were probed with a neurofilament heavy chain monoclonal antibody (Product # 711025, red) at a dilution of 1:250. Tissues were then incubated with Alexa Fluor 594 Goat anti-Mouse IgG (H+L) Secondary Antibody. Nuclei (blue) were stained with DAPI. Data courtesy of Min Sun Kim at Seoul National University, Korea.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Culture and identification of DPSCs. (A) Teeth of New Zealand white rabbits; (B) primary culture; (C) subculture; (D) immunohistochemical staining of vimentin; (E) alizarin red staining; (F) alkaline phosphatase staining; (G) immunofluorescence staining of NF200 (red); (H) immunofluorescence staining of Stro1 (green).