Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 105557 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 105557-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV9 | 105557-AAV9 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV(Search Vector Database)
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberUnknown
Gene/Insert
- Gene/Insert namemCerulean
- PromoterCB7
Cloning Information
- Cloning methodUnknown
Resource Information
- Supplemental Documents
- pENN.AAV.CB7.CI.mCerulean.WPRE.RBG (p3248).gb
- Terms and Licenses
- UBMTA
- Vanderbilt-Cerulean
- Industry Terms
- Not Available to Industry
Depositor Comments
Penn Vector Core number p3248
Information for AAV1 (Catalog # 105557-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pENN.AAV.CB7.CI.mCerulean.WPRE.RBG (#105557). In addition to the viral particles, you will also receive purified pENN.AAV.CB7.CI.mCerulean.WPRE.RBG plasmid DNA.
CB7-driven mCerulean expression. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ 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
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenemCerulean
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.
Information for AAV9 (Catalog # 105557-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pENN.AAV.CB7.CI.mCerulean.WPRE.RBG (#105557). In addition to the viral particles, you will also receive purified pENN.AAV.CB7.CI.mCerulean.WPRE.RBG plasmid DNA.
CB7-driven mCerulean expression. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ 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, AAV9 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV9
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenemCerulean
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.
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2. 检查流程:
(1) 检查时先让患者口服一粒碳14尿素胶囊。
(2) 静坐25分钟后,受试者直接向集气瓶内呼气,患者呼气后集气瓶中的液体由粉红色变成无色为止,或者患者持续呼气时间已经达到3分钟后即可停止呼气。
(3) 向集气瓶中加入4.5ml稀释闪烁液后,加盖旋紧,置于液闪仪中进行检测。
(4) 检查过程中患者应当保持安静,剧烈运动后血中的酸碱度变化可能影响同位素标记CO2的呼出,另外在患者呼气时应当嘱咐患者注意不要将集气瓶中的液体误吸入口腔。向左转|向右转
SAMPOSCPMELAPSEDSAMPLETOTALELAPSED
TIME(MIN)TIME(MIN)
1**-1827543.90.5250.892
765315.21.5251.892
729279.52.5002.867
这是使用同一个底物,但所测得值,不知道为什麽CPM值一直往下滑?
有几个问题想问问大家:
1.我看到相当多文献中,结束反应用的是抽滤,由于条件的限制,我们这里没有该装置,所以就取了少数文献中的方法,用12000G离心10min结束反应,然后沉淀TRIS-HCL洗涤3次。测量时在沉淀所在的EP管中加入闪烁液(该闪烁液的配方适用于样品含水量较少的情况),每管1ML。请问大家,这样结束反应行吗?沉淀是否应该烘干?
2.我用的液闪仪计数不是很稳定,如果进行校正,尤其是读数比较小的时候(如只有数百),以减少误差?
3.加入闪烁液后,是否应该避光放置一段时间后再测量?
谢谢!
碳14试剂检测方法
(1) 静坐25分钟后,受试者直接向集气瓶内呼气,患者呼气后集气瓶中的液体由粉红色变成无色为止,或者患者持续呼气时间已经达到3分钟后即可停止呼气。
(2) 向集气瓶中加入4.5ml稀释闪烁液后,加盖旋紧,置于液闪仪中进行检测。
(3) 检查过程中患者应当保持安静,剧烈运动后血中的酸碱度变化可能影响同位素标记CO2的呼出,另外在患者呼气时应当嘱咐患者注意不要将集气瓶中的液体误吸入口腔。
安全性
碳14尿素呼气试验应用于临床十几年,未见到明显的不良反应的报道。专业性评估报告证实碳14呼气试验对患者和操作人员的辐射危险可忽略不计,临床上可以安全使用。
十分感谢!!!
将反应所需的试剂与含有荧光素酶的细胞裂解液混合即会产生一种迅速衰减(在一秒钟内)的黄绿色闪光(发射峰560 nm),这种光信号可用配备了便于迅速混合反应物的自动注射装置的荧光检测仪(Luminometer)进行检测,也可用标准的液闪仪对光信号进行记录。当底物过量时,发光量的总值与样品的荧光酶活性成正比,因此,可对荧光素酶报告基因的转录进行间接估计。荧光素酶易被蛋白酶降解,在转染的哺乳动物细胞中的半衰期约为3 小时。荧光素酶报告系统为启动子活性的检测提供了一个敏感、快速、非放射性的检测方法。
荧光素酶报告基因检测试剂盒(Luciferase Reporter Gene Assay Kit),是以荧光素(luciferin)为底物来检测萤火虫荧光素酶(firefly luciferase)。荧光素酶可以催化luciferin氧化成oxyluciferin,在luciferin氧化的过程中,会发出生物荧光(bioluminescence)。然后可以通过荧光测定仪也称化学发光仪(luminometer)或液闪测定仪测定luciferin氧化过程中释放的生物荧光。
通过荧光素和荧光素酶这一生物发光体系,可以极其灵敏、高效地检测基因的表达。通常把感兴趣的基因转录的调控元件克隆在luciferase的上游或其他适当的地方,构建成报告基因质粒。然后转染细胞,适当刺激或处理后裂解细胞,测定荧光素酶活性。通过荧光素酶活性的高低判断刺激前后或不同刺激对感兴趣的调控元件的影响。展开
有几个问题想问问大家:
1.我看到相当多文献中,结束反应用的是抽滤,由于条件的限制,我们这里没有该装置,所以就取了少数文献中的方法,用12000G离心10min结束反应,然后沉淀TRIS-HCL洗涤3次。测量时在沉淀所在的EP管中加入闪烁液(该闪烁液的配方适用于样品含水量较少的情况),每管1ML。请问大家,这样结束反应行吗?沉淀是否应该烘干?
2.我用的液闪仪计数不是很稳定,如果进行校正,尤其是读数比较小的时候(如只有数百),以减少误差?
3.加入闪烁液后,是否应该避光放置一段时间后再测量?
谢谢!

