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StressMarq/Anti-Ubiquitin Antibody [FK2]/SMC-214D-A565/100-µg

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SMC-214D-A565

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Overview:

Product Name Ubiquitin Antibody

Description

Mouse Anti-Human Ubiquitin Monoclonal IgG1

Species Reactivity Species Independent

Applications WB, IHC, ICC/IF, IP, ELISA

Antibody Dilution WB (1:10000); optimal dilutions for assays should be determined by the user.

Host Species Mouse

Immunogen Species Human

Immunogen Ubiquitin conjugated lysozyme

Concentration 10 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	10mM phosphate buffer, 0.15M NaCl pH7.4, 0.1% sodium azide
Storage Temperature	-20ºC, 4ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Protein A purified
Clonality	Monoclonal
Clone Number	FK2
Isotype	IgG1
Specificity	Detects ubiquitinated proteins and ubiquitin chains. Does not detect free ubiquitin.
Cite This Product	StressMarq Biosciences Cat# SMC-214, RRID: AB_2699358
Certificate of Analysis	A 1:5000 dilution of SMC-214 was sufficient for detection of ubiquitin conjugates in 2.5 µg of HeLa cell lysate by electrochemiluminescence analysis using goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names	Polyubiquitin B Antibody, RPS27A Antibody, UBA52 Antibody, UBB Antibody, UBC Antibody, ubiquitin B Antibody
Research Areas	Cell Signaling, Post-translational Modifications, Ubiquitination
Cellular Localization	Cytoplasm, Nucleus
Accession Number	BAC56955.1
Gene ID	7314
Swiss Prot	P0CG47
Scientific Background	Ubiquitin is a small protein found ubiquitously in all tissue types and acts as a post translational modification. It can bind to its substrate either as a single ubiquitin molecule or in a chain. It is involved in many regulatory processes, which include proteasomal degradation, signal transduction, DNA repair, endocytosis and autophagy.
References	1. Chen J., & Chen Z. (2013). Curr Opin Immunol. (1): 4-12. 2. Shaid S., Brandts C., Serve H., & Dikic I. (2013). Cell Death Differ. 20(1): 21-30.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone FK2 (SMC-214). Tissue: Fibroblast cell line (NIH 3T3). Species: Mouse. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-214) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 60X.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone FK2 (SMC-214). Tissue: Fibroblast cell line (NIH 3T3). Species: Mouse. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-214) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 60X.

Product Citations (0)

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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

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

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有学预防的前辈吗?电泳后,正、负极槽内电泳缓冲液pH会怎样变化? ...123

曌猴速矩512021-08-05

将会降低

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

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。

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？

电泳缓冲液浓度对实验时间有影响吗123

才子不才00022017-10-02

电泳缓冲液浓度对实验时间有影响。（如下案例）试验在20-40mmol/L范围内考察了醋酸铵浓度对4中组分分离度的影响。结果发信啊4中组分的迁移时间随醋酸铵浓度的增大而延长，从而分离度得到改善。但是缓冲液浓度越大分析时间也越长，如图3-3所示。向左转|向右转

【求助】人血清脂蛋白琼脂糖凝胶电泳缓冲液蛋白质和糖学技术...123

czp862021-07-22

本人对电泳了解不多，现在有个问题向大家求教；我做的是人血清脂蛋白琼脂糖凝胶电泳，我看文献上基本上是写的是用Tris-巴比妥缓冲液，但现在巴比妥和巴比妥钠很贵，用量很大，而且还买不到！我想问下可不可用其他什么缓冲液代替一下？请帮忙，谢谢大家了

【求助】为什么我的SDSPAGE电泳时电泳缓冲液变浑浊呢?123

戀█重量█p82017-10-02

原因可能如下：
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

第二向sdspage,电泳缓冲液可以多次使用吗? 生物科学 ...123

chromosomedx2007-08-12

在《分子克隆》上看见检测微量的DNA可以使用先电泳，再用含EB的电泳缓冲液染色30至45分钟。
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^_^

SDSPAGE电泳缓冲液的上槽液与下槽液有何区别? 123

轩子TA01942017-10-02

1。阳极缓冲液 ( 10 × ) ：121.14g Tris 溶于400ml 蒸馏水，用 1.0mol/L HCl 调至 pH8.9 ,定容至
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。

关于电泳缓冲液,奇怪又好笑的问题蛋白质和糖学专业医生社区...123

woxinyijiu2021-07-30

问个关于电泳缓冲液的问题，如果用TBE作为缓冲液来做胶，然后用TAE来作为缓冲液来跑胶，或者是倒过来，电泳结果会不会有什么变化？

tbe电泳缓冲液配方_完美作业网_www.wanmeila.com123

214302552017-10-02

没什么实质性的差别。制胶、作为电泳缓冲液都可以。不过作为电泳缓冲液时，0.5×TBE更容易变热。

如果你是制胶或者做电泳缓冲液的话，不用灭菌。

【求助】几种蛋白电泳缓冲液的异同以及PVDF转膜过程中缓冲液要...123

zcxy43162021-07-24

目前常用的SDS-PAGE电泳缓冲液有：
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！