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StressMarq/Anti-DYKDDDDK Tag Antibody (Equivalent to Sigma’s Anti-FLAG® M2 Antibody)/SPC-720D-A488/100-µg

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StressMarq/Anti-DYKDDDDK Tag Antibody (Equivalent to Sigma’s Anti-FLAG® M2 Antibody)/SPC-720D-A488/100-µg

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StressMarq

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SPC-720D-A488

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Overview:

Product Name DYKDDDDK Tag Antibody (Equivalent to Sigma’s Anti-FLAG® M2 Antibody)

Description

Rabbit Anti- DYKDDDDK Tag Polyclonal

Species Reactivity Species Independent

Applications WB

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

Host Species Rabbit

Immunogen Synthetic peptide corresponding to the DYKDDDDK Tag, conjugated to Keyhole Limpet Haemocyanin (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 pH 7.4, 50% glycerol, 0.09% Sodium Azide
Storage Temperature	-20ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Peptide Affinity Purified
Clonality	Polyclonal
Specificity	Binds to DYKDDDDK (FLAG tag) conjugated proteins. Recongizes the same epitope as Sigma's Anti-FLAG® antibodies.
Cite This Product	StressMarq Biosciences Cat# SPC-720, RRID: AB_2706131
Certificate of Analysis	A 1:1000 dilution of SPC-720 was sufficient for detection of DYKDDDDK Tag in 15 µg of DYKDDDDK (FLAG) Positive Control Lysate cell lysates by ECL immunoblot analysis using goat anti-rabbit IgG:HRP as the secondary antibody.

Biological Description

Alternative Names	DDDDK epitope tag antibody, DDDK antibody, ddk antibody, DYKDDDDK antibody, DYKDDDDK epitope tag antibody, DYKDDDDK tag antibody, ECS epitope tag antibody, ECS tag antibody, Enterokinase Cleavage Site epitope tag antibody, Enterokinase Cleavage Site tag antibody, FLAG antibody, FLAG tag antibody
Research Areas	Epitope Tags, Tags and Cell Markers

Product Images

<p>Dot Blot analysis using Rabbit Anti-DYKDDDDK Tag Polyclonal Antibody (SPC-720). Tissue: DYKDDDDK (FLAG) Positive Control Lysate. Primary Antibody: Rabbit Anti-DYKDDDDK Tag Polyclonal Antibody (SPC-720) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Rabbit HRP:IgG at 1:3000 for 1 hour at RT. The quantities of protein spotted on each panel are as shown.</p>

Dot Blot analysis using Rabbit Anti-DYKDDDDK Tag Polyclonal Antibody (SPC-720). Tissue: DYKDDDDK (FLAG) Positive Control Lysate. Primary Antibody: Rabbit Anti-DYKDDDDK Tag Polyclonal Antibody (SPC-720) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Rabbit HRP:IgG at 1:3000 for 1 hour at RT. The quantities of protein spotted on each panel are as shown.

Product Citations (0)

Currently there are no citations for this product.

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

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

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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

【求助】WB上样缓冲液加加多少是怎么确定的? 蛋白质和糖学...123

你大爷RgZ2017-10-02

上样缓冲液也有多种，一般是80ul的样本加20ul的上样缓冲液，这样基本上上样50ug的体积在10ul左右。
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。

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

【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

请教:2*SDS上样缓冲液的配法蛋白质和糖学论坛123

小2b1雿231k2017-10-02

2× SDS-PAGE 上样缓冲液：
0.5 M tris-HCl (pH 6.8) 20％
丙三醇 20％
20％SDS 20％
0.1％溴酚兰 5％
2-巯基乙醇 10％
双蒸水 25％

不调PH值，PH在配0.5 M tris-HCl 时已调，缓冲液呈蓝紫色。

【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？

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