Overview:
| Product Name | DMPO Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Description | Mouse Anti-DMPO Monoclonal IgG1 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Species Reactivity | Species Independent | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Applications | WB, IHC, ICC/IF, IP, ELISA, AM | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Antibody Dilution | WB (1:1000), ICC/IF (1:100), ELISA (1:100), IP (25µg); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Host Species | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen | 5,5-dimethyl-2-(8-octanoic acid)-1-pyrrolone-N-oxide conjugated to Ovalbumin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Concentration | 0.48 mg/ml, 1 mg/ml | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Conjugates |
Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
StreptavidinProperties:
Streptavidin Datasheet Biotin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| R-PE (R-Phycoerythrin) | ||
Overview:
R-PE Datasheet | ![]() | Optical Properties: λex = 565 nm λem = 575 nm εmax = 2.0×106 Φf = 0.84 Brightness = 1.68 x 103 Laser = 488 to 561 nm Filter set = TRITC |
Properties
| Storage Buffer | PBS pH7.4, 50% glycerol, 0.09% sodium azide |
| Storage Temperature | -20ºC |
| Shipping Temperature | Blue Ice or 4ºC |
| Purification | Protein G Purified |
| Clonality | Monoclonal |
| Clone Number | N1664A |
| Isotype | IgG1 |
| Specificity | Recognizes DMPO, DMPO-octanoic acid, DMPO-protein adducts and DMPO-DNA adducts. Does not cross react with non-adducted proteins or DNA. |
| Cite This Product | StressMarq Biosciences Cat# SMC-189, RRID: AB_10703686 |
| Certificate of Analysis | A 1:1000 dilution of SMC-189 was sufficient to detect the DMPO nitrone adducts of metmyoglobin when loaded at 100 ng/lane by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody. |
Biological Description
| Alternative Names | 5,5-dimethyl-2-(8-octanoic acid)-1-pyrroline N oxide Antibody, DMPO nitrone adduct Antibody, 55 dimethyl 1 pyrroline N oxide nitrone adduct antibody |
| Research Areas | Cancer, Cell Signaling, Oxidative Stress |
| Scientific Background | The formation of free radicals and other highly reactive oxygen species has been implicated in the pathogenesis of many disease states (1). The ability to identify these species is crucial, and spin trapping has accomplished this goal. DMPO (5,5-dimethyl-1-pyrroline N-oxide) is one of the least toxic to cells and animals, and possesses convenient pharmacokinetics (uptake, distribution, metabolism and excretion) in biological systems (2-6). Recent studies have determined that nitric oxide may substantially affect the quantitative determination of DMPO adducts, and therefore extra caution is required when studying generation of these species in the presence of nitric oxide or its radicals (1). DMPO adducts can be generated with protein and DNA radicals (7). |
| References |
1. Reszka K.J., et al. (2006) Nitric Oxide 15: 133-141. 2. Ramirez D.C., Gomez-Mejiba S.E., and Mason R.P. (2007) Nat Protoc. 2(3): 512-522. 3. Khan N., et al. (2003) Free Radic. Biol. Med 34:1473–1481. 4. Haseloff R.F., et al. (1997) FEBS Lett 418:73–75. 5. Schaefer C.F., Janzen E.G., West M.S., Poyer J.L., and Kosanke S.D. (1996) Free Radic. Biol. Med 21:427–436. 6. Anzai K., et al. (2003) Arch. Biochem. Biophys 415:251–256. 7. Free Radic Biol Med. (2009) April 1; 46(7): 853–865. doi:10.1016/j.freeradbiomed.2008.12.020. 8. Chatterjee S., et al. (2009) Free Radic. Med.and Biol. 46: 454-461. |
Product Images
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-DMPO Monoclonal Antibody, Clone N1664A (SMC-189). Tissue: macrophage cell line (Raw 264.7). Species: Mouse. Primary Antibody: Mouse Anti-DMPO Monoclonal Antibody (SMC-189) at 1:100. Secondary Antibody: Alexa Fluor 488 Goat Anti-Mouse (green) at 1:1000. Counterstain: DAPI (blue) nuclear stain.
Western Blot analysis of Human HL 60 clone 15 eosinophils lysates showing detection of DMPO protein using Mouse Anti-DMPO Monoclonal Antibody, Clone N1664A (SMC-189). Primary Antibody: Mouse Anti-DMPO Monoclonal Antibody (SMC-189) at 1:200.
Product Citations (2)
Other Citations
Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.
Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.
PubMed ID: N/A Reactivity Human Applications: Antibody Microarray
Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.
Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.
PubMed ID: N/A Reactivity Mouse Applications: Antibody Microarray
| ATTO 488 | ||
Overview:
ATTO 488 Datasheet | ![]() | Optical Properties: λex = 501 nm λem = 523 nm εmax = 9.0×104 Φf = 0.80 τfl = 4.1 ns Brightness = 72 Laser = 488 nm Filter set = FITC |
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② 应取pH8.0,这样可使核苷酸带较多负电荷,利于吸附于阴离子交换树脂柱。虽然pH 11.4时核苷酸带有更多的负电荷,但pH过高对分离不利。
③ 当不考虑树脂的非极性吸附时,根据核苷酸负电荷的多少来决定洗脱速度,则洗脱顺序为CMP>AMP> GMP > UMP,但实际上核苷酸和聚苯乙烯阴离子交换树脂之间存在着非极性吸附,嘌呤碱基的非极性吸附是嘧啶碱基的3倍。静电吸附与非极性吸附共同作用的结果使洗脱顺序为:CMP> AMP > UMP >GMP。
1. Buffer中离子浓度过大,甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高,也是一个原因
我使用的是1%琼脂糖凝胶,电泳缓冲液为0.5×TBE。
请问各位大虾,在不影响实验结果的前提下,这样的含EB的电泳缓冲液可以反复使用多少次?
谢谢^_^
500ml
2。阴极缓冲液 ( 10 × ) :将 60.55g Tris ,89.58g Tricine 及 5g SDS 溶于400ml 蒸馏水中,加水至终体积为500ml
使用时稀释至1×的缓冲液,电泳槽内槽加入阴极电泳缓冲液,外槽加入阳极电泳缓冲液。形成Trcine-SDS-PAGE电泳系统。
如果你是制胶或者做电泳缓冲液的话,不用灭菌。
Tris-Glycine/SDS;
MOPS/SDS;
Bis-Glycine/SDS等不同的缓冲液、预混液等。
不知道这几种缓冲液有什么异同呢?
比如如果是Invitrogen的预制胶-预混液电泳系统,每种预制胶有适配缓冲液,但是如果用其他的缓冲液似乎也可以得到不错的结果,那么是否说明电泳缓冲液只要满足了缓冲ph范围,其他的不是特别重要呢?
此外,附加提问是,PVDF膜在转膜的时候,转膜缓冲液中不需要加入甲醇,那么是加入甲醇转膜效果好还是不加好呢?
谢谢!















