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StressMarq/Anti-DUX4 Antibody [P2B1]/SMC-192D-ALP/100-µg

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SMC-192D-ALP

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Overview:

Product Name DUX4 Antibody

Description

Mouse Anti-Human DUX4 Monoclonal IgG1

Species Reactivity Human, Mouse

Applications WB, IHC, ICC/IF

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

Host Species Mouse

Immunogen Species Human

Immunogen C-terminal 76 amino acids of DUX4 with glutathione-s-transferase (gst) tag

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 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	P2B1
Isotype	IgG1
Specificity	Detects ~45kDa. No cross-reactivity with DUX4c.
Cite This Product	StressMarq Biosciences Cat# SMC-192, RRID: AB_10964369
Certificate of Analysis	1 µg/ml of SMC-192 was sufficient for detection of DUX4 in 20 µg of HeLa cell lysate by ECL immunoblot analysis using goat anti-mouse IgG: HRP as teh secondary.

Biological Description

Alternative Names	Double homeobox 4 Antibody, Double homeobox 10 Antibody, DUX10 Antibody, Double homeobox protein 4/10 Antibody
Research Areas	Cell Signaling
Cellular Localization	Nucleus
Accession Number	NP_149418.3
Gene ID	22947
Swiss Prot	F5GZ66
Scientific Background	DUX4, or double homeobox4, is a human protein that is a transcriptional activator of paired-like homeodomain transcription factor 1 (1). Clinically it is a facioscapulohumeral muscular dystrophy candidate gene that appears to have a toxic gain of function (2-4). In FSHD individuals, the expression of the full-length DUX4 transcript is not completely suppressed in skeletal muscle and possibly other differentiated tissues (5).
References	1. Entrez Gene: “Dux4 Double Homeobox, 4” 2. Dixit M., et al., (2007) Proc. Natl Acad Sci. USA. 104(46): 18157-18162. 3. Kowaljow V., et al. (2007) Neuromuscl Disord. 17(8): 611-623. 4. Lemmers R., et al. (2010) Science Express. 329(5999): 1650-1653. 5. Snider L., et al. (2010) PLoS Genetics. 6:1-14.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-DUX4 Monoclonal Antibody, Clone P2B1 (SMC-192). Tissue: C2C12 myoblast cells. Species: Mouse. Primary Antibody: Mouse Anti-DUX4 Monoclonal Antibody (SMC-192) at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green). Counterstain: DAPI (blue) nuclear stain.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-DUX4 Monoclonal Antibody, Clone P2B1 (SMC-192). Tissue: C2C12 myoblast cells. Species: Mouse. Primary Antibody: Mouse Anti-DUX4 Monoclonal Antibody (SMC-192) at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green). Counterstain: DAPI (blue) nuclear stain.

<p>Western Blot analysis of Mouse C2C12 cell lysate showing detection of DUX4 protein using Mouse Anti-DUX4 Monoclonal Antibody, Clone P2B1 (SMC-192). Primary Antibody: Mouse Anti-DUX4 Monoclonal Antibody (SMC-192) at 1:1000. Cells transfected with pCS2+DUX4 which, contains an additional upstream start site.</p>

Western Blot analysis of Mouse C2C12 cell lysate showing detection of DUX4 protein using Mouse Anti-DUX4 Monoclonal Antibody, Clone P2B1 (SMC-192). Primary Antibody: Mouse Anti-DUX4 Monoclonal Antibody (SMC-192) at 1:1000. Cells transfected with pCS2+DUX4 which, contains an additional upstream start site.

Product Citations (0)

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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

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芷玥丹青2017-10-02

上样缓冲液中甘油与溴酚蓝的左右分别是
① 上样缓冲液中包括甘油,溴酚蓝,SDS等,甘油主要作用是防止样品漂浮出点样孔,溴酚蓝作为电泳指示剂,用来观察电泳进度,SDS用于一些核酸结合蛋白的变性解离.
②核酸染料与凝胶中核酸结合,以便于在紫外光下显示核酸条带.

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？

4×和5×的上样缓冲液区别| 吉至试剂123

zmp189198708772021-07-31

4×和5×的上样缓冲液区别

【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

【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

【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

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

【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？

dna提取过程加入缓冲液b为什么会产生白色沉淀 123

txstbbm2010-03-27

找公司合成了一个30AA的阳离子多肽（纯度90%），加水溶解至6ng/μl,加入2x上样缓冲液后，立即出现沉淀，加热至99度10分钟仍不溶解，离心取上清电泳，蛋白Marker条带清晰，样品无条带形成，在预期位置的上方10~15kd处出现弥散状着色区，请问：
1）为何出现沉淀，怎样才能避免沉淀的出现？
2）为何没有预期大小的3.5kd条带出现？10~15kd处可能是什么？是不是分子聚合？若是分子聚合，怎样解聚？

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。

dna上样缓冲液和rna上样缓冲液可以通用么_点样缓冲液_缓冲液_...123

小P孩01752021-08-06

dna上样缓冲液和rna上样缓冲液可以通用。
因为DNA和RNA链上的碱基都基本一致。而这两种上样缓冲液中含有的染料是二甲苯青和溴酚蓝，都可以与其碱基结合，跑出条带。
但是，如果使用DNA上样缓冲液，里面会夹杂一些RNA酶，这些酶有可能会影响到实验结果。使用前需要将RNA酶处理掉。