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

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SPC-1294D

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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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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。