TDC2 antibody (C-Term)

Catalog No. ABIN4889606

Product Details anti-TDC2 Antibody

Target: TDC2 (Tyrosine Decarboxylase 2 (TDC2))

Binding Specificity: C-Term

Reactivity: Drosophila melanogaster

Host: Rabbit

Clonality: Polyclonal

Application: ELISA, Western Blotting (WB), Immunohistochemistry (IHC)

Specificity: Reacts with Drosophila melanogaster 72 kDa Tdc2 protein

Purification: Purified (protein A)

Immunogen: Synthetic peptide derived from C-terminal part of Drosophila Tdc2 protein.

Application Details

Application Notes:

Working dilution: Optimal dilutions should be determined by the end user.
The following are guidelines only:
IHC(1:200 - 1:1000) WB(1:200 - 1:2000)

Restrictions: For Research Use only

Independent Validation (IF)

Validation #100812 (Immunofluorescence)

by: Department of Entomology, University of California, Riverside

No.: #100812

Date: 05/26/2017

Antigen: Tdc2

Lot Number: 13B1

Method validated: Immunofluorescence

Positive Control:

D. melanogaster octopaminergic neurons labeled with Tdc2-Gal4 of the abdominal nerve to the ovary

Notes:

Passed. ABIN809182 labels octopaminergic neurons specifically and with no background.

Validation Images

Immunostaining of D. melanogaster octopaminergic ovarian nerves expressing GFP (A), immunostaining of expressed GFP (green) (B), and of tdc2 with ABIN4889606 (red) (C). The three channels are merged in D.

Full Methods

Primary Antibody: ABIN4889606

Secondary Antibody: goat anti-rabbit AF542 conjugated antibody (Life Technologies)

Full Protocol:

• Dissect ovaries of D. melanogaster ETHR-Gal4/UAS-MCD8-GFP expressing GFP in octopaminergic neurons in cold Schneider’s Insect Medium (S2; Sigma Aldrich, S01416).
• Transfer tissue to 2ml protein LoBind tubes (Eppendorf, 022431102) filled with S2 containing 2% paraformaldehyde (PFA) at RT.
• Fix tissue for 55min at RT while nutating.
• Wash tissue 4x 10min with 1.75ml PBS containing 0.5% Triton X-100 (PBST).
• Remove PBST and add 200µl 5% goat serum (GS; Thermo Fisher Scientific, 16210064) in PBST per tube.
• Incubate 1.5h at RT on a rotator.
• Remove blocking solution.
• Incubate with primary
• • rabbit anti-Tdc2 antibody (Tyrosine Decarboxylase 2) (C-Term) (antibodies-online, ABIN4889606, lot 13B1) diluted 1:200 in blocking solution.
  • mouse anti-GFP (Thermo Fisher Scientific) diluted 1:500 in blocking solution.
• Incubate for 4h at RT followed by 36-48h at 4°C on a rotator.
• Rinse tissue with 1.75ml PBST. Allow the tissue to settle to the bottom before removing the liquid.
• Wash tissue 3x 30min with 1.75ml PBST.
• Incubate with secondary 200µl secondary goat anti-rabbit AF542 conjugated antibody (Life Technologies) and goat anti-mouse AF488 conjugated antibody (Life Technologies, A11034) diluted 1:500 in blocking solution containing 0.5mg/ml DAPI.
• Incubate for 4h at RT followed by 72h at 4°C on a rotator.
• Rinse tissue with 1.75ml PBST. Allow the tissue to settle to the bottom before removing the liquid.
• Wash tissue 3x 30min with 1.75ml PBST.
• Add 1.75ml PBST containing 4% PFA at RT.
• Fix tissue for 5h at RT while nutating.
• Rinse tissue with 1.75ml PBST. Allow the tissue to settle to the bottom before removing the liquid.
• Wash tissue 4x 15min with 1.75ml PBST.
• Mount tissue on a poly-L-lysine (Sigma Aldrich, P1524-25MG) coated cover glass.
• Dehydrate tissue by covering the cover glass for 10min each with 30%, 50%, 75%, 95%, 100%, 100%, and 100% EtOH.
• Clearing using by covering the cover glass 3x 5min with xylene.
• Add 7 drops of dibutyl phthalate in xylene (DPX) on top of the mounted tissue.
• Seat cover glass face down gently onto a prepared slide with spacers.
• Let the slide dry for 48h at RT in a hood before viewing.

Experimental Notes:

• Staining of ETHR-Gal4/UAS-MCD8-GFP expressing GFP in octopaminergic neurons was performed as previously described.

• ABIN4889606 worked fantastically. It labeled octopaminergic neurons specifically and with no background. Pictures are below. The well-characterized neurons labeled with Tdc2-Gal4 of the abdominal nerve to the ovary described in Middleton et al. (2006) overlapped with ABIN4889606. Staining with ABIN4889606 was stronger than Gal4 labeling.

Handling

Format: Lyophilized

Reconstitution: Must be reconstituted in distilled water.

Concentration: 1 mg/mL

Buffer: Tris 0,1M, glycine 0,1M, sucrose 2 %

Storage: 4 °C/-20 °C

Storage Comment: Lyophilized powder stable for a minimum of 2 years at -20°C. Store reconstituted antibodies at +4°C. For extended periods store in aliquots at -20°C. Antibodies are guaranteed for 6 month from date of receipt.

Expiry Date: 24 months

Target Details for TDC2

Target: TDC2 (Tyrosine Decarboxylase 2 (TDC2))

Alternative Name: Tyrosine Decarboxylase 2

Synonyms: CG30446 antibody, CG3686 antibody, Dmel\CG30446 antibody, TDC2 antibody, dTdc2 antibody, tdc antibody, tdc2 antibody, Tyrosine decarboxylase 2 antibody, Tdc2 antibody

Background: Enzyme involved in tyrosine metabolism. Use Pyridoxal phosphate as cofactor.

Gene ID: 246620

UniProt: A1Z6N4