PA1-18371

antibody from Invitrogen Antibodies

Targeting: BDNF

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

PA1-18371

Provider

Invitrogen Antibodies

Product name

BDNF Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Reconstitute in 100 µL of sterile water. Centrifuge to remove any insoluble material. After reconstitution keep aliquots at -20 °C for a higher stability, and at 4 °C with an appropriate antibacterial agent. Glycerol (1:1) may be added for an additional stability. Avoid repetitive freeze/thaw cycles.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-BDNF Polyclonal Antibody (Product # PA1-18371) and a 34kDa band corresponding to Brain-derived neurotrophic factor was observed across the panel tested except for Mouse liver and spleen which are reported to be negative. Whole cell extract and tissue extract (30 µg lysate) of U-87 MG (Lane 1), IMR-32 (Lane 2), SW480 (Lane 3), SiHa (Lane 4), HeLa (Lane 5), Mouse Brain (Lane 6), Mouse Liver (Lane 7), Mouse Spleen (Lane 8), Rat Brain (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Brain-derived neurotrophic factor was performed using 70% confluent log phase SW480 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with BDNF Polyclonal Antibody (Product # PA1-18371) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:3000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.