TDC2 抗体 (Tyrosine Decarboxylase 2) (C-Term)

Details for Product anti-TDC2 Antibody No. ABIN4889606, 供应商: Log in to see
抗原
  • CG30446
  • CG3686
  • Dmel\CG30446
  • TDC2
  • dTdc2
  • tdc
  • tdc2
  • Tyrosine decarboxylase 2
  • Tdc2
抗原表位
C-Term
1
适用
Fruit Fly (Drosophila melanogaster)
1
宿主

1
克隆类型
多克隆
应用范围
ELISA, Immunohistochemistry (IHC), Western Blotting (WB)
1
1
选项
Supplier
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Supplier Product No.
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'独立验证'标志
抗原 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.

Primary Antibody ABIN4889606
Secondary Antibody goat anti-rabbit AF542 conjugated antibody (Life Technologies)
实验流程
  • 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.

Validation Images
Immunofluorescence validation image for anti-Tyrosine Decarboxylase 2 (TDC2) (C-Term) antibody (ABIN4889606)

Immunostaining of D. melanogaster octopaminergic ovarian nerves expressing ...

免疫原 Synthetic peptide derived from C-terminal part of Drosophila Tdc2 protein.
特异性 Reacts with Drosophila melanogaster 72 kDa Tdc2 protein
纯化方法 Purified (protein A)
别名 Tyrosine Decarboxylase 2
背景 Enzyme involved in tyrosine metabolism. Use Pyridoxal phosphate as cofactor.
基因ID 246620
UniProt A1Z6N4
应用备注

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)

限制 仅限研究用
状态 Lyophilized
溶解方式 Must be reconstituted in distilled water.
浓度 1 mg/mL
缓冲液 Tris 0,1M, glycine 0,1M, sucrose 2 %
储存条件 4 °C/-20 °C
储存方法 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.
有效期 24 months
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