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StressMarq/Anti-DMPO Antibody [N1664A]/SMC-189D/100-µg

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SMC-189D

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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

PerCP

Overview:

Peridinin-Chlorophyll-Protein Complex
Small phycobiliprotein
Isolated from red algae
Large stokes shift (195 nm)
Molecular Weight: 35 kDa

PerCP Datasheet

PerCP Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 482 nm

λ_em = 677 nm

ε_max = 1.96 x 10⁶

Laser = 488 nm

PE/ATTO 594

PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.

Overview:

High fluorescence yield
High photostability
Very hydrophilic
Excellent solubility in water
Very little aggregation

PE/ATTO 594 Datasheet

PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra

Optical Properties:

λ_ex = 535 nm

λ_em = 627 nm

Laser = 488 to 561 nm

FITC (Fluorescein)

Overview:

Excellent fluorescence quantum yield
High rate of photobleaching
Good solubility in water
Broad emission spectrum
pH dependent spectra
Molecular formula: C₂₀H₁₂O₅
Molar mass: 332.3 g/mol

FITC-Fluorescent-conjugate

FITC Fluorescein Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 494 nm

λ_em = 520 nm

ε_max = 7.3×10⁴

Φ_f = 0.92

τ_fl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

ATTO 700

Overview:

High fluorescence yield
Excellent thermal and photostability
Quenched by electron donors
Very hydrophilic
Good solubility in polar solvents
Zwitterionic dye
Molar Mass: 575 g/mol

ATTO 700 Datasheet

ATTO 700 Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 700 nm

λ_em = 719 nm

ε_max = 1.25×10⁵

Φ_f = 0.25

τ_fl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

ATTO 680

Overview:

High fluorescence yield
Excellent thermal and photostability
Quenched by electron donors
Very hydrophilic
Good solubility in polar solvents
Zwitterionic dye
Molar Mass: 631 g/mol

ATTO 680 Datasheet

ATTO 680 Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 680 nm

λ_em = 700 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

ATTO 655

Overview:

High fluorescence yield
High thermal and photostability
Excellent ozone resistance
Quenched by electron donors
Very hydrophilic
Good solubility in polar solvents
Zwitterionic dye
Molar Mass: 634 g/mol

ATTO 655 Datasheet

ATTO 655 Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 663 nm

λ_em = 684 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

ATTO 633

Overview:

High fluorescence yield
High thermal and photostability
Moderately hydrophilic
Good solubility in polar solvents
Stable at pH 4 – 11
Cationic dye, perchlorate salt
Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

ATTO 633 Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 629 nm

λ_em = 657 nm

ε_max = 1.3×10⁵

Φ_f = 0.64

τ_fl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

ATTO 594

Overview:

High fluorescence yield
High photostability
Very hydrophilic
Excellent solubility in water
Very little aggregation
New dye with net charge of -1
Molar Mass: 1137 g/mol

ATTO 594 Datasheet

ATTO 594 Fluorophore Excitation and Emission Spectrum

Optical Properties:

λ_ex = 601 nm

λ_em = 627 nm

ε_max = 1.2×10⁵

Φ_f = 0.85

τ_fl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

ATTO 565

Overview:

High fluorescence yield
High thermal and photostability
Good solubility in polar solvents
Excellent solubility in water
Very little aggregation
Rhodamine dye derivative
Molar Mass: 611 g/mol

ATTO 565 Datasheet

ATTO 565 Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 563 nm

λ_em = 592 nm

ε_max = 1.2×10⁵

Φ_f = 0.9

τ_fl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

ATTO 488

Overview:

High fluorescence yield
High photostability
Very hydrophilic
Excellent solubility in water
Very little aggregation
New dye with net charge of -1
Molar Mass: 804 g/mol

ATTO 488 Datasheet

ATTO 488 Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 501 nm

λ_em = 523 nm

ε_max = 9.0×10⁴

Φ_f = 0.80

τ_fl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

ATTO 390

Overview:

High fluorescence yield
Large Stokes-shift (89 nm)
Good photostability
Moderately hydrophilic
Good solubility in polar solvents
Coumarin derivate, uncharged
Low molar mass: 343.42 g/mol

ATTO 390 Datasheet

ATTO 390 Fluorescent Dye Excitation and Emission Spectra

Optical Properties:

λ_ex = 390 nm

λ_em = 479 nm

ε_max = 2.4×10⁴

Φ_f = 0.90

τ_fl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

APC (Allophycocyanin)

Overview:

High quantum yield
Large phycobiliprotein
6 chromophores per molecule
Isolated from red algae
Molecular Weight: 105 kDa

APC Datasheet

APC Fluorophore Absorption and Emission Spectrum

Optical Properties:

λ_ex = 650 nm

λ_em = 660 nm

ε_max = 7.0×10⁵

Φ_f = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

Streptavidin

Properties:

Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
Molecular weight: 53 kDa
Formula: C₁₀H₁₆N₂O₃S
Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

Biotin

Properties:

Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
Molar mass: 244.31 g/mol
Formula: C₁₀H₁₆N₂O₃S
Applications: Western blot, immunohistochemistry, and ELISA

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

Enzymatic activity is used to amplify weak signals and increase visibility of a target
Readily combines with hydrogen peroxide (H₂O₂) to form HRP-H₂O₂ complex which can oxidize various hydrogen donors
Catalyzes the conversion of:
- Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
- Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
- Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
High turnover rate enables rapid generation of a strong signal
44 kDa glycoprotein
Extinction coefficient: 100 (403 nm)
Applications: Western blot, immunohistochemistry, and ELISA

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
Catalyzes the conversion of:
- Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
- Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
Molecular weight: 140 kDa
Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

R-PE (R-Phycoerythrin)

Overview:

Broad excitation spectrum
High quantum yield
Photostable
Member of the phycobiliprotein family
Isolated from red algae
Excellent solubility in water
Molecular Weight: 250 kDa

R-PE Datasheet

R-PE Fluorophore Excitation and Emission Spectra

Optical Properties:

λ_ex = 565 nm

λ_em = 575 nm

ε_max = 2.0×10⁶

Φ_f = 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

<p>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.</p>

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.

<p>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.</p>

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

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SDSPAGE电泳相关溶液的配置123

qinhuan04252021-08-01

最近刚开始跑SDS-PAGE，很奇怪，加样后，Marker和样本停在加样孔不往下面跑！且跑电泳时，电压（80V）一直上不去，后测电泳液PH值，不到8.0，这是主要原因吗？

如需调PH，是用NaOH吗？

电泳分离四种核苷酸时,通常将缓冲液调到什么pH?此时它们是向哪极移动...123

阿瑟6102021-08-09

① 电泳分离4种核苷酸时应取pH3.5 的缓冲液，在该pH时，这4种单核苷酸之间所带负电荷差异较大，它们都向正极移动，但移动的速度不同，依次为：UMP>GMP>AMP>CMP；
　　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。

蛋白Mops电泳缓冲液是怎么回事儿– 960问答123

浮1502017-10-02

MOPS电泳缓冲液、胚胎干细胞培养生长因子、动物肝细胞分离培养试剂盒、结晶紫细胞群落染色试剂盒、MFN-2蛋白表达检测试剂盒、血红细胞溶解液、磷酸缓冲盐溶液、Hanks平衡盐粉剂、胰蛋白酶溶液、氯霉素溶液等产品由上海哈灵生物有限公司提供

【求助/交流】蛋白电泳缓冲液分子生物综合其他论坛...123

coolaber2021-07-20

北京COOLABER隆重推出PBS、TAE、TBE、蛋白电泳缓冲液独立包装预混干粉试剂，配缓冲液像冲咖啡一样简单，您只需要准备容器和水，就一切OK啦！什么计算配比，什么称重，什么调PH值，都去他的！联系电话：010-8244451715132878855400-878-6800QQ:1002073414李云云

电泳时,缓冲液为弱酸性或中性,DNA为什么会带负电?123

猴琢型182017-10-02

DNA分子在高于其等电点的pH溶液中中带负电,因为所说的等电点是指蛋白质或两性电解质所带净电荷为零时溶液的pH,此时蛋白质或两性电解质在电场中的迁移率为零.当外界溶液的pH大于两性离子的等电点值,两性离子释放质子带负电；当外界溶液的pH小于两性离子的等电点值,两性离子质子化带正电.

电泳缓冲液TAE与TBE,哪个更好? – BioEngX123

coolaber2021-08-03

【求助】跑SDSPAGE的电泳缓冲液需要什么水来配制经验共享 ...123

小聪用的03832017-10-02

跑SDS-PAGE的电泳缓冲液需要什么水来配制，我以前都用一馏水配制，但是有时配制好的电泳缓冲液比较混倒入电泳槽中，都看不清楚琼脂糖封底的下面有没有气泡。最近实验室有人用给细胞配液用的二馏水配制缓冲液，很清澈，跑的电流电压好像也没受什么影响啊，不是说电泳时需要有离子的吗，二馏水里基本是什么都没有了啊。另外，电泳时电流与什么有关，我跑电泳时同样的电压但是电流都不相同，而且现在电流没有以前大了电泳时间也延长很多

【求助】电泳缓冲液多长时间更换一次啊 PCR技术讨论版论坛123

老丽丽2021-08-10

如题啊如题，多久时间换一次好些呢？？:O):O):O):O)

关于电泳缓冲液,奇怪又好笑的问题蛋白质和糖学技术讨论版...123

灬丨牛牛丨灬2021-07-31

笔者小白，做WB时发现了一个奇怪的问题：

电泳缓冲液购买Solarbio的成品（5X），后稀释成1X工作液。

电泳条件：80V120V（注：内外槽每次用的都是新配液体，未使用回收后液体，且泳槽无漏液现象）

电流仪器：Bio-Rad

奇怪的事情在于：当我设定恒定电压为80V跑的时候，查看电流显示为35mA，且电流数值随着时间变化不断降低，半小时后约降至23mA。

请教大神们，之前有没有遇到这样的问题？这倒底是什么原因造成的，应该如何改善？

【求助】为什么我的SDSPAGE电泳时电泳缓冲液变浑浊呢?123