GDNF antibody (AA 121-211)

Catalog No. ABIN736536

This anti-GDNF antibody is a Rabbit Polyclonal antibody detecting GDNF in WB, ELISA, FACS, IF (cc), IF (p), IHC (p) and IHC (fro). Suitable for Human, Mouse, Rat, Cow and Rabbit. Independently validated for use in Immunofluorescence. This Primary Antibody has been cited in 1 publication.

Quick Overview for GDNF antibody (AA 121-211) (ABIN736536)

Target: GDNF (Glial Cell Line Derived Neurotrophic Factor (GDNF))

Reactivity: Human, Mouse, Rat, Cow, Rabbit

Host: Rabbit

Clonality: Polyclonal

Conjugate: This GDNF antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Product Details anti-GDNF Antibody

Binding Specificity: AA 121-211

Cross-Reactivity: Cow, Human, Mouse, Rabbit, Rat

Predicted Reactivity: Dog,Cow,Pig,Horse,Chicken

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human GDNF

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IF)

Validation #102003 (Immunofluorescence)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #102003

Date: 02/12/2018

Antigen: GDNF

Lot Number: 9F01M8

Method validated: Immunofluorescence

Positive Control: mouse hippocampus and putamen

Negative Control: no primary control

Notes:

ABIN736536 works in IF albeit with some background staining that we have been unable to eliminate although using different dilutions to increase the signal-to-noise ratio.

Validation Images

IF staining with ABIN736536 of mouse dorsal root ganglion (DRG, A), the dorsal horn of the spinal cord (B), and the hippocampus CA1 (C) and Putamen (D).

Full Methods

Primary Antibody: ABIN736536

Secondary Antibody: anti-rabbit AF488 conjugated antibody (Life Technologies)

Full Protocol:

• Perfuse mouse with 4% paraformaldehyde in 0.1M phosphate buffer (PB) pH7.4.
• Post-fix spinal cord and brain blocks in the same fixative for additional 2h at RT.
• Wash spinal cord and brain blocks several times with PBS.
• Cut blocks with a vibratome (Leica, VT1000 S) into 70µm thick transverse sections.
• Cut dorsal root ganglions (DRGs) with a cryostat into 17µm thick sections after cryoprotection and glass mounting.
• Block free floating vibratome and glass mounted cryostat sections with blocking solution (0.01M PBS5% Normal Goat Serum (NGS; Sigma, G9023, lot SLBV1396), 0.1% Triton X-100 (BioRad, 161-0407, lot 00583) for 1h at RT.
• Incubate sections with primary rabbit anti-GDNF antibody (antibodies-online, ABIN736536, lot 9F01M8) diluted 1:100 in blocking solution ON at RT.
• Wash sections 4x for 5min with 0.01M PBS.
• Incubate sections with secondary goat anti-rabbit AF488 conjugated antibody (Life Technologies) diluted 1:500 in PBS for 1h at RT.
• Wash sections 4x for 5min with 0.01M PBS.
• Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).

Experimental Notes:

• Different dilutions were also tested (1:200, 1:500, 1:2000) with or without Triton-X but the 1:100 dilution and the use of Triton-X in the blocking solution gave the best results.

• Staining is mainly present in the cytoplasm of small-size neurons in the DRG, consistent with what has been previously described. ABIN736536 stains fibers in superficial laminae of the dorsal horn of the spinal cord. In the hippocampus staining is present in the CA1 both in the pyramidal neurons (cell body and dendrites) and in individual interneurons. In the putamen ABIN736536 stains neuron cell bodies and some dendrites.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-GDNF antibody (ABIN736536)

Zhang, Cai, Song, Dong, Hou, Lv: "Normalization of ventral tegmental area structure following acupuncture in a rat model of heroin relapse." in: Neural regeneration research, Vol. 9, Issue 3, pp. 301-7, (2014) (PubMed).

Target Details for GDNF

Target: GDNF (Glial Cell Line Derived Neurotrophic Factor (GDNF))

Alternative Name: GDNF

Background:

Synonyms: ATF1, ATF2, HSCR3, HFB1-GDNF, Glial cell line-derived neurotrophic factor, hGDNF, Astrocyte-derived trophic factor, ATF, GDNF

Background: Neurotrophic factor that enhances survival and morphological differentiation of dopaminergic neurons and increases their high-affinity dopamine uptake.

Gene ID: 2668

UniProt: P39905

Pathways: RTK Signaling, Synaptic Membrane, Tube Formation, Autophagy, Smooth Muscle Cell Migration