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StressMarq/Anti-DNMT1 Antibody [11H8]/SMC-231D-PCP/100-µg

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StressMarq/Anti-DNMT1 Antibody [11H8]/SMC-231D-PCP/100-µg

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StressMarq

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SMC-231D-PCP

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Overview:

Product Name DNMT1 Antibody

Description

Mouse Anti-Human DNMT1 Monoclonal IgG1

Species Reactivity Human, Mouse

Applications WB, ICC/IF

Antibody Dilution WB (1:1000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.

Host Species Mouse

Immunogen Species Human

Immunogen Raised against a synthetic peptide corresponding to amino acids 637-650 of human DNMT1

Concentration 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, 0.05% BSA, 0.05% sodium azide, 50% glycerol
Storage Temperature	-20ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Protein G Purified
Clonality	Monoclonal
Clone Number	11H8
Isotype	IgG1
Specificity	Detects ~180kDa. It will cross-react with mouse DNMT1.
Cite This Product	StressMarq Biosciences Cat# SMC-231, RRID: AB_2699448
Certificate of Analysis	2 µg/ml of SMC-231 was sufficient for detection of Dnmt1 in 10 µg of mouse ES cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names	DNA Mtase Antibody, DNMT Antibody, MCMT Antibody, DNA methyltransferase 1 Antibody, AIM Antibody, CXXC9 Antibody, DNMT Antibody, DNA (cytosine-5)-methyltransferase 1 Antibody, CXXC-type zinc finger protein 9 Antibody, DNA methyltransferase HsaI Antibody
Research Areas	Cell Signaling, Organelle Markers, Post-translational Modifications
Cellular Localization	Nucleus
Accession Number	NP_001370
Gene ID	1786
Swiss Prot	P26358
Scientific Background	Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA may be linked with development of cancer. To date, 3 families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of this gene causes global demethylation of genomic DNA and embryonic lethality. Dnmt2 is expressed at low levels in adult tissues and its inactivation does not affect DNA methylation or maintenance of methylation. The Dnmt3 family members, Dnmt3a and Dnmt3b, are strongly expressed in ES cells but their expression is down regulated in differentiating ES cells and is low in adult somatic tissue. Dnmt1 co-purifies with the retinoblastoma (Rb) tumour suppressor gene product, E2F1, and HDAC1. Dnmt1 also cooperates with Rb to repress transcription from promoters containing E2F-binding sites suggesting a link between DNA methylation, histone deacetylase and sequence-specific DNA binding activity, as well as a growth-regulatory pathway that is disrupted in nearly all cancer cells (1-6).
References	1. Bestor T., et al.(1988) J.Mol. Biol. 203: 971-983. 2. Yen R.W., Vertino P.M., Nelkin B.D., et al. (1992) Nucl. Acids Res. 20: 2287-2291. 3. Xie S., et al. (1999) Gene 236: 87-95. 4. Okano M., Bell D.W., Haber D.A. and Li E. (1999) Cell 99: 247-257. 5. Reik W. et al. (1999) J. Nat. Genet 23: 380-382. 6. Robertson K.D., et al. (2000) Nat Genet 25(3): 338-342.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 11H8 (SMC-231). Tissue: Colon carcinoma cell line (RKO). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody (SMC-231) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60 min at RT, 5 min at RT. Localization: Nucleus. Magnification: 60X. (A) DAPI nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) DNMT1. (D) Composite.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-DNMT1 Monoclonal Antibody, Clone 11H8 (SMC-231). Tissue: Colon carcinoma cell line (RKO). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-DNMT1 Monoclonal Antibody (SMC-231) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60 min at RT, 5 min at RT. Localization: Nucleus. Magnification: 60X. (A) DAPI nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) DNMT1. (D) Composite.

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PerCP

Overview:

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

PerCP Datasheet

Optical Properties:

λ_ex = 482 nm

λ_em = 677 nm

ε_max = 1.96 x 10⁶

Laser = 488 nm

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【求助】上样缓冲液的问题经验共享分析测试百科 123

冰凝果842021-07-22

上样缓冲液是4×，上样体积是24微升，则上样缓冲液应该是6微升。如果不按这个比例，多加或者少加有什么影响呢？谢谢

【求助】蛋白质变性时是不是必须加上样缓冲液经验共享分析测试...123

黎约风骚e32021-07-23

是

双向电泳样品缓冲液选用的基本原则123

神伿2017-10-02

电泳缓冲液的主要作用是使胶的导电性和体系的一致,这样跑出的条带才回一致；
加样缓冲液的主要作用是使PCR产物与其混合,
使DNA沉于加样孔的底部,防止DNA跑出来.

5X的SDSPAGE蛋白上样缓冲液配方有谁知道4X的也可以 123

黎约全球の15962017-10-02

total:10ml 4度下保存：
1mol/l Tirs-HCl(PH8.8) 0.6ml
10%SDS 2ml
50%甘油 5ml
2-巯基乙醇 0.5ml
1%溴酚兰 1ml
水 0.9ml
另外：SDS不好溶解建议加热溶解,如果要做非还原SDS-PAGE请将2-巯基乙醇 0.5ml换成超纯水即可

上样缓冲液中溴酚蓝和蔗糖溶液的作用 123

4671607912021-07-20

作为指示剂，因为溴酚蓝呈蓝色，而蛋白质是白色，在SDS-凝胶中不明显，且溴酚蓝的相对分子量比蛋白质(绝大部分)小，电泳时速度比蛋白质稍快，因此当溴酚蓝到达电泳槽底部(可看蓝色调带)，则电泳结束。

fot tsukuardgothic std m字体|fot tsukuardgothic std m字体下载...123

浊影2021-07-27

我做的是明胶酶谱实验，开始用以前师姐剩下的还原型上样缓冲液（成分不详）上样组织蛋白酶，电泳后洗脱掉SDS，置入反应液中，酶可以在相应位置降解明胶，这次换了新的上样缓冲液，就是含巯基乙醇的，可是电泳后酶却不能降解明胶了，不知道这个巯基乙醇打开蛋白质的结构后，对酶的活性有没有不可逆的影响，如果有的话，是不是要换一种上样缓冲液了？谢谢

请问western的5X上样缓冲液具体用量? 分子生物综合其他...123

吴文昊冖2021-07-30

Tris-甘氨酸电泳缓冲液:30.3gTris,188g甘氨酸,10gSDS,用蒸馏水溶解至1000ml,得0.25mol/L Tris-1.92mol/L甘氨酸电极缓冲液.临用前稀释10倍.

【求助】western blot上样缓冲液蛋白质和糖学论坛123

woailucy2021-08-02

实验室的一份protocol 上写配上样缓冲液的时候要加溴酚蓝bromophenol blue，但没写加的量是多少。实验室只有粉末状的溴酚蓝。请问大神们如何加，加多少啊？需要先把溴酚蓝配制成溶液吗？还是直接加进去？

做WB,蛋白加上样缓冲液煮过之后,又超声了一次,对蛋白有影响吗...123

会下蛋的鱼2021-08-03

提了蛋白，加上样缓冲液煮过，但发现还是有很多胶状的沉淀，比较黏稠，堵枪头。于是自作主张又超声了一次，结果跑了一次，内参都没有。煮过之后的蛋白不能超声么？

超详细!WB实验蛋白浓度定量与上样体积计算Protocol分享!_样品123

打鼓ikCO2021-08-07

凝胶电泳是为了分离不同物理性质（如大小、形状、等电点等）的分子。加入缓冲溶液是为了调节合适的离子浓度和PH值，保持分离物质分子构象不变。缓冲液在电泳过程中的一个作用是维持合适的pH。电泳时正极与负极都会发生电解反应，正极发生的是氧化反应（4OH--4e->2H2O+O2)，负极发生的是还原反应（4H++4e->2H2)，长时间的电泳将使正极变酸，负极变碱。一个好的缓冲系统应有较强的缓冲能力，是溶液两极的pH保持基本不变。

电泳缓冲液的另一个作用是使溶液具有一定的导电性，以利于DNA分子的迁移，例如，一般电泳缓冲液中应含有0.01-0.04mol/L的Na+离子，Na+离子的浓度太低时电泳速度变慢；太高时就会造成过大的电流使胶发热甚至熔化。

电泳缓冲液还有一个组分是EDTA，加入浓度为1-2mmol/L，目的是螯合Mg2+ 等离子，防止电泳时激活 DNA酶，此外还可防止Mg2+离子与核酸生成沉淀。

用于dna琼脂糖凝胶电泳的上样缓冲液及其制备方法123

fsj19872021-08-04

同仁好，我想请教一下大家，6Xloddingbuffer1%琼脂糖凝胶电泳TBE缓冲液，5ul
DNA样品和1ul上样缓冲液怎么就是跑不出主带啊？都是弥散带，给点建议我哪里有问题啊？

5*蛋白质上样缓冲液怎么配? 分子生物学术科研...123

mollysjy2017-10-02

2ul 5×上样缓冲液中添加3ul水，混匀，即得2×上样缓冲液