
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 104496 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 104496-AAV1 | 100 µL at titer ≥ 7×10¹² vg/mLand Plasmid.More Information | Add to Cart | |||
AAV Retrograde | 104496-AAVrg | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backboneAAV-CAG-FLEX(Search Vector Database)
- Backbone manufacturerScott Sternson
- Backbone sizew/o insert(bp)5027
- Total vector size (bp)6380
- Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature30°C
- Growth Strain(s)NEB Stable
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namejGCaMP7f
- Alt nameGCaMP3-T302L R303P A317L D380Y
- Alt nameGCaMP3 variant 921
- Alt nameJanelia GCaMP7
- SpeciesR. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
- Insert Size (bp)1353
- PromoterCAG
- Tag/ Fusion Protein
- T7 epitope, Xpress tag, 6xHis
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBsmBI(destroyed during cloning)
- 3′ cloning siteBsmBI(destroyed during cloning)
- 5′ sequencing primerGGTTCGGCTTCTGGCGTGTGACC (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
Information for AAV1 (Catalog # 104496-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE (#104496). In addition to the viral particles, you will also receive purified pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE plasmid DNA.
CAG-driven, Cre-dependent jGCaMP7f calcium sensor. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
Information for AAV Retrograde (Catalog # 104496-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE (#104496). In addition to the viral particles, you will also receive purified pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE plasmid DNA.
CAG-driven, Cre-dependent jGCaMP7f calcium sensor. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
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1.若都为正常,那么并未有太大问题,只要定期复查就可以了。
2.如果TGAb、TMAb、甲状腺功能均高,那么就是桥本氏甲状腺炎并甲亢,需要抗甲亢治疗。
3.如果TGAb、TMAb高,甲状腺功能下降,那么就是桥本氏甲状腺炎并甲减,需要进行甲减治疗。
4.如果TGAb、TMAb高,甲状腺功能正常,那么就是桥本氏甲状腺炎,无需特殊治疗,只要定期复查甲状腺功能就可能以了,不过这种情况有可能以后演变成甲减或者甲亢。
1.若都为正常,那么并未有太大问题,只要定期复查就可以了。
2.如果TGAb、TMAb、甲状腺功能均高,那么就是桥本氏甲状腺炎并甲亢,需要抗甲亢治疗,那么,要注意戒碘饮食,避免吃一些含碘丰富的食物,如海产品等。
3.如果TGAb、TMAb高,甲状腺功能下降,那么就是桥本氏甲状腺炎并甲减,需要进行甲减治疗,饮食方面可以增加含碘丰富的食物摄入。
4.如果TGAb、TMAb高,甲状腺功能正常,那么就是桥本氏甲状腺炎,无需特殊治疗,只要定期复查甲状腺功能就可能以了,不过这种情况有可能以后演变成甲减或者甲亢。
1.若都为正常,那么并未有太大问题,只要定期复查就可以了。
2.如果TGAb、TMAb、甲状腺功能均高,那么就是桥本氏甲状腺炎并甲亢,需要抗甲亢治疗。
3.如果TGAb、TMAb高,甲状腺功能下降,那么就是桥本氏甲状腺炎并甲减,需要进行甲减治疗。
4.如果TGAb、TMAb高,甲状腺功能正常,那么就是桥本氏甲状腺炎,无需特殊治疗,只要定期复查甲状腺功能就可能以了,不过这种情况有可能以后演变成甲减或者甲亢。
请问这个图是怎么做出来的?
辣根过氧化物酶+DAB显色和免疫荧光染色可以同时在一张脑片里面做吗?
具体是先DAB显色还是先染荧光一抗二抗呢,具体实验步骤是怎样的呢?
第一张图是往血管里打入辣根过氧化物酶,后DAB显色,DIC成像
第二张图是萤光图

