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StressMarq/Anti-DNMT1 Antibody [11H8]/SMC-231S/12-µg

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SMC-231S

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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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ysh14152017-10-02

最好有图片说明，谢谢

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是

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最好是测浓度，一般不超过30-40ug，上样缓冲液一般4×或者5×，这个没太大区别。

【求助】做western时,蛋白浓度太低怎么办蛋白质和糖学123

山楂冰糖大白梨2017-07-22

实验小白刚刚起步，一般采用等质量上样的方法，但我算的上样量是没加缓冲液之前的，加完缓冲液蛋白样品浓度会变么？师姐说不会变，就按照算好的上样量加就行。

前几天提的样按50ug算每个样本的上样量也只有上5ul左右，按师姐说的就那么上了，结果发完光只有内参有条带，目标蛋白一点也没有，但以前做过一次是出过一点趋势的，而且这次整个过程感觉做的很仔细，发完光结果也很干净整齐，所以我在想是不是跟上样量有关系？上样缓冲液加入之后会使样本浓度稀释五倍么，这样的话上样量就得乘5。

变性前后加上样缓冲液的区别是什么呢？哪一种会改变蛋白浓度从而改变上样量呢？

【电泳实验中什么叫上样缓冲液?WESTERNBLOT中需要进行电泳,其具体...123

犬夜叉IQK2017-10-02

蛋白上样缓冲液
蛋白电泳上样缓冲液包括2%SDS , 0.1%溴酚蓝 10%甘油,SDS是保证样品中的所有蛋白带电荷一致，减少电荷对电泳结果的影响。还原剂是让二硫键处于断开状态，保证蛋白分子的线性.
溴酚蓝作用是指示上样蛋白的跑胶时标记的，甘油是增加上样重量的，以免漂浮，煮沸是使蛋白变性的永久保存

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？

SDS电泳溶液的配置123

2021-08-05

sds电泳上样缓冲液如何配置

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

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.

【求助】新手请问2×上样缓冲液,DTT怎么配制?什么时候加?? 蛋白...123

清风烧饼782017-10-02

（以下内容仅供参考）
2乘SDS-PAGE上样缓冲液（20ml体系）
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水定容至20ml
4℃冰箱保存，可以不分装，我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的

4×蛋白上样缓冲液使用说明123

songjunlai19922016-03-22

请问有人知道蛋白上样缓冲液应该怎么配制吗？请赐教