TDC2 抗体 (C-Term)
Quick Overview for TDC2 抗体 (C-Term) (ABIN4889606)
抗原
适用
宿主
克隆类型
应用范围
- 
    - 
                                            抗原表位
- C-Term
- 
                                            特异性
- Reacts with Drosophila melanogaster 72 kDa Tdc2 protein
- 
                                            纯化方法
- Purified (protein A)
- 
                                            免疫原
- Synthetic peptide derived from C-terminal part of Drosophila Tdc2 protein.
 
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    - 
                                            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
 
- 
                                            
- 
        - by
- Department of Entomology, University of California, Riverside
- No.
- #100812
- 日期
- 2017.05.26
- 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 
- Negative Control
- Notes
- Passed. ABIN809182 labels octopaminergic neurons specifically and with no background. 
- 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. 
 
 生效 #100812 (Immunofluorescence)![成功验证 '独立验证'标志]()  ![成功验证 '独立验证'标志]() Validation ImagesFull Methods Validation ImagesFull Methods
- 
    - 
                                            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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- 
    - TDC2 (Tyrosine Decarboxylase 2 (TDC2))
- 
                                            别名
- Tyrosine Decarboxylase 2
- 
                                            背景
- Enzyme involved in tyrosine metabolism. Use Pyridoxal phosphate as cofactor.
- 
                                            基因ID
- 246620
- 
                                            UniProt
- A1Z6N4
 抗原
- 
                    
 
                                                 (1 validation)
 (1 validation) 
                 
                                     
                                     
                                    