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Overview:

Product Name Ubiquitin Antibody

Description

Rabbit Anti-Bovine Ubiquitin Polyclonal

Species Reactivity Dog, Human, Monkey, Mouse, Rat, African clawed frog (Xenopus laevis), Bovine, Chicken, Fish, Fruit Fly (Drosophila melanogaster), Guinea Pig (Cavia porcellus), Hamster, Pig, Rabbit, Rainbow Trout (Oncorhynchus mykiss), Sheep, Yeast

Applications WB, IHC, ICC/IF, IP

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

Host Species Rabbit

Immunogen Species Bovine

Immunogen Native bovine Ubiquitin, conjugated to KLH

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	Peptide Affinity Purified
Clonality	Polyclonal
Specificity	Detects ~10kDa. It also recognizes ubiquinated proteins.
Cite This Product	StressMarq Biosciences Cat# SPC-119, RRID: AB_2285248
Certificate of Analysis	A 1:1000 dilution of SPC-119 was sufficient for detection of free ubiquitin in 15 µg of HeLa lysate by ECL immunoblot analysis using Donkey anti-rabbit 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, Cell membrane, Nucleus
Accession Number	NP_776558.1
Gene ID	281370
Swiss Prot	P0CG53
Scientific Background	Ubiquitin is a small protein that occurs in all eukaryotic cells. The ubiquitin protein itself consists of 76 amino acids and has a molecular mass of about 8.5 kDa. Key features include its C-terminal tail and the 7 Lys residues. It is highly conserved among eukaryotic species: Human and yeast ubiquitin share 96% sequence identity (1). The main function of Ubiquitin is to clear abnormal, foreign and improperly folded proteins by targeting them for degradation by the 26S proteosome (2). Ubiquitination represents an essential cellular process affected by a multi-enzyme cascade involving classes of enzymes known as ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s or Ubcs) and ubiquitin-protein ligases (E3s). Ubiquitin is activated in a two-step reaction by an E1 ubiquitin-activating enzyme in a process requiring ATP as an energy source. The initial step involves production of an ubiquitin-adenylate intermediate. The second step transfers ubiquitin to the E1 active site cysteine residue, with release of AMP. This step results in a thioester linkage between the C-terminal carboxyl group of ubiquitin and the E1 cysteine sulfhydryl group. The third step is a transfer of ubiquitin from E1 to the active site cysteine of a ubiquitin-conjugating enzyme E2 via a trans(thio)esterification reaction. And the final step of the ubiquitylation cascade creates an isopeptide bond between a lysine of the target protein and the C-terminal glycine of ubiquitin. In general, this step requires the activity of one of the hundreds of E3 ubiquitin-protein ligases (often termed simply ubiquitin ligase). E3 enzymes function as the substrate recognition modules of the system and are capable of interaction with both E2 and substrate(3, 4). Ubiquitination also participates in the internalization and degradation of plasma membrane proteins such as some of the TCR subunits while still ER-membrane associated (5). Ubiquitin also plays a role in regulating signal transduction cascades through the elimination inhibitory proteins, such as IκBα and p27 (6).
References	1. Wilkinson K.D. (1995) Annu. Rev. Nutr. 15:161-189. 2. Smalle J., Vierstra R.D. (2004) Anu Rev Plant Biol. 55: 555-590. 3. Bonifacino J.S., et al. (1998) Annu Rev Cell Dev Biol. 14: 19-57. 4. Boston Biochem: "Ubiquitin Proteasome Pathway Overview” http://www.bostonbiochem.com/upp.php 5. Yang M., et al. (1998) J Exp Med. 187: 1835-1846. 6. Chen Z.J., et al. (1996) Cell 84: 853-862.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite. Heat Shocked at 42°C for 1h.</p>

Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite. Heat Shocked at 42°C for 1h.

<p>Western blot analysis of Human HEK93 lysates showing detection of Ubiquitin protein using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:1000.</p>

Western blot analysis of Human HEK93 lysates showing detection of Ubiquitin protein using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:1000.

<p>Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative Solution. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Cytoplasm.</p>

Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative Solution. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Cytoplasm.

<p>Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT.</p>

Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT.

<p>Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:200 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite.</p>

Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:200 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite.

Product Citations (1)

Western Blot

EMG1 harbouring the Bowen-Conradi syndrome D86G substitution localizes normally but has reduced stability.

Armistead, J. and Triggs-Raine, B. (2013) Ribosomes: Molecular Structure, Role in Biological Functions and Implications for Genetic Diseases. ISBN: 978-1-62417-698-2.

PubMed ID: N/A Reactivity Human Applications: Western Blot

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

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

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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？

TE缓冲液 123

☆你大爷☆pgyn2021-07-29

TE缓冲液是Tris +EDTA缓冲液，这种缓冲液我们一般用作溶解剂或保持剂，主要是调控PH，TAE是一种电泳缓冲液，主要用于DNA分子的电泳。 TE组成浓度：10mM Tris-HCl 1mM EDTA PH=8.0 配制量：500ml 配制方法：量取下列溶液于500ml烧杯中 1M Tris-HCl Buffer PH=8.0 5ml 0.5M EDTA PH=8.0 1ml 向烧杯中加入约400mldd H2O均匀混合；将溶液定容到500ml后，高温高压灭菌；室温保存.
TAE是使用最广泛的缓冲系统。其特点是超螺旋在其中电泳时更符合实际相对分子质量（TBE中电泳时测出的相对分子质量会大于实际分子质量），且双链线状DNA在其中的迁移率较其他两种缓冲液快约10%，电泳大于13kb的片段时用TAE缓冲液将取得更好的分离效果，此外，回收DNA片段时也易用TAE缓冲系统进行电泳。TAE的缺点是缓冲容量小，长时间电泳（如过夜）不可选用，除非有循环装置使两极的缓冲液得到交换。 50×TAE Buffer 配制方法： 1. 称量Tris 242g，Na2EDTA.2H2O 37.2g 于1L烧杯中； 2.向烧杯中加入约800ml去离子水，充分搅拌均匀； 3加入57.1ml的冰乙酸，充分溶解； 4.加去离子水定容至1L后，室温保存。向左转|向右转

【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

RNA电泳为什么要用MOPS缓冲液123

嗳情啖了﹎4142021-08-08

电泳缓冲液还有一个组分是EDTA（乙二胺四乙酸），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。
此外，我们常常用“电泳法”判断液体的性质，是胶体还是溶液。在高中化学中我们就用过这种方法判断给定的液体是否为胶体。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、

【评测】High Five昆虫细胞培养基123

angel_lei2021-07-25

急需TricineSDS-PAGE电泳中阴阳电泳缓冲液的配置！
谢谢各位了。
我找了几篇文章，但是写的都很含糊，不知道有没有找到具体的数据的？

为什么电泳时采用PH8.6的缓冲液 123

cjh一一2017-10-02

电泳缓冲液的ph为8.6时为什么会向正极移动

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？

电泳法分离蛋白质时,缓冲液的离子强度的一般要求是A.0.01～0.05B....123

想自由zJ72021-07-28

离子强度是表示溶液中电荷数量的一种量度，等于溶液中各种离子的摩尔浓度与其价数平方之积总和的一半。
低离子强度时，迁移率快。但离子强度过低，缓冲液的缓冲容量小，不易维持pH恒定。高离子强度时迁移率慢，因此为了获得更快的反应速度，在缓冲容量允许的范围内，离子强度应该尽可能小。