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StressMarq/Anti-Doublecortin Antibody/SPC-1294D-RPE/100-µl

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StressMarq

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SPC-1294D-RPE

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Overview:

Product Name Doublecortin Antibody

Description

Rabbit Anti-Human Doublecortin Polyclonal

Species Reactivity Human, Mouse, Rat

Applications WB, ELISA

Antibody Dilution WB (1:1000), ELISA (1:10000); optimal dilutions for assays should be determined by the user.

Host Species Rabbit

Immunogen Species Human

Immunogen Synthesized unphosphorylated peptide derived from human Doublecortin around the phosphorylation site of serine 376 (P-T-SP-P-G).

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, 150mM NaCl, 0.02% sodium azide
Storage Temperature	-20ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Affinity Purified
Clonality	Polyclonal
Isotype	IgG
Specificity	Detects ~45kDa, endogenous levels of total Doublecortin protein.
Cite This Product	StressMarq Biosciences Cat# SPC-1294, RRID: AB_2713097
Certificate of Analysis	A 1:1000 dilution of SPC-1294 was sufficient for detection of Doublecortin in 10 µg of Jurkat cell lysates by ECL immunoblot analysis using Goat Anti-Rabbit IgG:HRP as the secondary antibody.

Biological Description

Alternative Names	DBCN Antibody, Dbct Antibody, DC Antibody, DCX Antibody, DCX_HUMAN Antibody, Doublecortex Antibody, Doublin Antibody, FLJ51296 Antibody, Lis X Antibody, Lis-X Antibody, Lissencephalin X Antibody, Lissencephalin-X Antibody, Lissencephaly X linked Antibody, Lissencephaly X linked doublecortin Antibody, LISX Antibody, Neuronal migration protein doublecortin Antibody, OTTHUMP00000023859 Antibody, OTTHUMP00000023860 Antibody, OTTHUMP00000216315 Antibody, OTTHUMP00000216316 Antibody, SCLH Antibody, XLIS Antibody
Research Areas	Cell Markers, Cell Structure, Neuroscience
Cellular Localization	Cytoplasm, Cell projection, Cytoskeleton, Cytosol
Accession Number	NP_000546.2
Gene ID	1641
Swiss Prot	O43602
Scientific Background	Neuronal migration protein doublecortin is a protein that in humans is encoded by the DCX gene, also known as doublin or lissencephalin-X. Doublecortin (DCX) is a microtubule-associated protein expressed almost exclusively in immature neurons. Neuronal precursors begin to express DCX shortly after exiting the cell cycle, and continue to express DCX for 2-3 weeks as the cells mature into neurons. It seems to be required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. May act by competing with the putative neuronal protein kinase DCAMKL1 in binding to a target protein. May in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway.
References	1. Oomen C.A. et al. (2009) PLoS ONE 4 (1): e3675. 2. Brown J.P. et al. (2003) J. Comp. Neurol. 467 (1): 1–10. 3. Couillard-Despres S. et al. (2005) Eur. J. Neurosci. 21 (1): 1–14.

Product Images

<p>Western blot analysis of Human Jurkat cell lysates showing detection of ~40-45kDa Doublecortin protein using Rabbit Anti-Doublecortin Polyclonal Antibody (SPC-1294). Lane 1: Human Jurkat. Lane 2: Human Jurkat treated with the immunizing peptide. Primary Antibody: Rabbit Anti-Doublecortin Polyclonal Antibody (SPC-1294) at 1:1000. Predicted/Observed Size: ~40-45kDa.</p>

Western blot analysis of Human Jurkat cell lysates showing detection of ~40-45kDa Doublecortin protein using Rabbit Anti-Doublecortin Polyclonal Antibody (SPC-1294). Lane 1: Human Jurkat. Lane 2: Human Jurkat treated with the immunizing peptide. Primary Antibody: Rabbit Anti-Doublecortin Polyclonal Antibody (SPC-1294) at 1:1000. Predicted/Observed Size: ~40-45kDa.

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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

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

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【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。

上样缓冲液中溴酚蓝和蔗糖溶液的作用 123

4671607912021-07-20

作为指示剂，因为溴酚蓝呈蓝色，而蛋白质是白色，在SDS-凝胶中不明显，且溴酚蓝的相对分子量比蛋白质(绝大部分)小，电泳时速度比蛋白质稍快，因此当溴酚蓝到达电泳槽底部(可看蓝色调带)，则电泳结束。

DNA电泳相关试剂及缓冲液的配制123

怕解题2021-07-28

50×TAE Buffer 配制方法：
1。称量氨基丁三醇 242g，Na2EDTA.2H2O 37.2g 于1L烧杯中；
2。向烧杯中加入约600ml去离子水，充分搅拌均匀；
3。加入57.1ml的冰乙酸，充分溶解；
4。用NaOH调pH至8.3，加去离子水定容至1L后，室温保存。
使用时稀释50倍即1×TAE Buffer
10×TBE Buffer配制方法：
1。称量氨基丁三醇 108g，Na2EDTA.2H2O 7.44g，硼酸55g 于1L烧杯中；
2。加入约700ml去离子水，搅拌均匀；
3。用NaOH调pH至8.3，加去离子水定容至1L后，室温保存。
使用时稀释10倍即1×TBE Buffer

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】为什么蛋白与上样缓冲液混合后形成沉淀,急!_问答详情_...123

tqch_31520042009-07-30

蛋白质（经硫酸铵和等电点沉淀后用1MKCL复溶）一加2x或6X上样缓冲液就产生沉淀，试问何故？怎样解决？
经搜索发现类似的问题，但人家的裂解液中含盐酸胍，而我的溶液中是不含含盐酸胍啊。
请做蛋白质的行家帮忙解答，谢谢。

怎样2X的SDSPAGE蛋白上样缓冲液浓缩了?如题.还有2X和5X指的...123

彩虹兔兔55562017-10-02

不太好操作，里面有小分子的SDS、β巯基乙醇，较难浓缩，不是有4X的买么，还可以自己配啊，网上都有现成的配方，溴酚蓝、SDS、β巯基乙醇。。。

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

【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】蛋白加上样缓冲液煮后有沉淀生成蛋白质和糖学技术讨论...123

三毛爱看书2021-07-26

提取蛋白，western上样前，加上样缓冲液煮，煮之前是澄清透明的，煮完就有挺多的沉淀生成，不知道什么原因，怎么解决，求帮忙！

WB试剂配方123

异鸣23692021-07-25

10 mmol/L Tris-HCl(pH 8.0) 1 mmol/L EDTA(pH 8.0) 因为含有以上两种物质，所以称为TE。
配制分三步： 1) 1 M Tris-HCl (pH 8.0) 50 ml的配制：称取Tris碱6.06 g，加超纯水40 ml溶解，滴加浓HCl约2.1 ml调pH至8.0，定容至50 ml。

【求助】上样缓冲液中的溴酚蓝、甘油影响BCA定蛋白吗? 蛋白质...123

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.

SDS电泳溶液的配置123

2021-08-05

sds电泳上样缓冲液如何配置