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StressMarq/Anti-Cav1.2 Antibody [S57-46]/SMC-300D-FITC/100-µg

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StressMarq/Anti-Cav1.2 Antibody [S57-46]/SMC-300D-FITC/100-µg

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StressMarq

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SMC-300D-FITC

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Overview:

Product Name Cav1.2 Antibody

Description

Mouse Anti-Rabbit Cav1.2 Monoclonal IgG1

Species Reactivity Human, Mouse, Rat, Hamster

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

Antibody Dilution WB (1:1000), IHC (1:1000), ICC/IF (1:100), IP (1:200); optimal dilutions for assays should be determined by the user.

Host Species Mouse

Immunogen Species Rabbit

Immunogen Fusion protein amino acids 1507-1733 (intracellular carboxyl terminus) of rabbit Cav1.2

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, 0.1% sodium azide
Storage Temperature	-20ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Protein G Purified
Clonality	Monoclonal
Clone Number	S57-46
Isotype	IgG1
Specificity	Detects ~240kDa (varies with cell background due to glycosylation).
Cite This Product	StressMarq Biosciences Cat# SMC-300, RRID: AB_2069099
Certificate of Analysis	1 µg/ml of SMC-300 was sufficient for detection of Cav1.2 in 10 µg of rat brain lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names	CACH3 Antibody, CACN4 Antibody, CACNA 1D Antibody, CACNL1A2 Antibody, voltage dependent L type calcium channel subunit alpha 1D Antibody, alpha-1 subunit voltage-dependent calcium channel Antibody, calcium channel voltage-dependent L type alpha 1C subunit1 Antibody, voltage-gated calcium channel alpha subunit Cav1.2 Antibody, calcium channel L type Antibody, alpha 1 polypeptide isoform 1 cardic muscle Antibody, calcium channel cardic dihydropyridine-sensitive alpha-1 subunit Antibody, voltage-gated L-type calcium channel Cav1.2 alpha 1 subunit splice variant 10 Antibody, DHPR alpha-1 subunit Antibody, Voltage-gated calcium channel subunit alpha Cav1.2 Antibody, Calcium channel L type alpha-1 polypeptide isoform 1 cardiac muscle Antibody
Research Areas	Cancer, Calcium Channels, Cardiovascular System, Cell Signaling, Heart, Ion Channels, Neuroscience, Voltage-Gated Calcium Channels
Cellular Localization	Cell membrane, Membrane
Accession Number	NP_001129994.1
Gene ID	100144322
Swiss Prot	P15381
Scientific Background	Cav1.2 is a cardiac L-type calcium channel, and is important for excitation and contraction of the heart (1). It may be associated with a variant of Long QT syndrome called Timothy's syndrome (2, 3) and also with Brugada syndrome. Some references also suggest it is related to bipolar disease as well (3).
References	1. Splawski I., et al. (2004) Cell. 119 (1): 19-31. 2. Krey J.F., and Dolmetsch R. (2009) Biophysical. 96 (3): 221a-222a. 3. Crotti L., Celano G., Dagradi F. and Schwartz P.J. (2008) Orphanet J Rare Disease 3:18.

Product Images

<p>Immunohistochemistry analysis using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: hippocampus. Species: Human. Fixation: 10% formalin. Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.</p>

Immunohistochemistry analysis using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: hippocampus. Species: Human. Fixation: 10% formalin. Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

<p>Immunoprecipitation analysis using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: INS-1E cells. Species: Rat. Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:200. Courtesy of: Merrie Mosedale.</p>

Immunoprecipitation analysis using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: INS-1E cells. Species: Rat. Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:200. Courtesy of: Merrie Mosedale.

<p>Western Blot analysis of Hamster T-CHO cell lysate showing detection of CaV1.2 Calcium Channel protein using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:1000.</p>

Western Blot analysis of Hamster T-CHO cell lysate showing detection of CaV1.2 Calcium Channel protein using Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody, Clone S57-46 (SMC-300). Primary Antibody: Mouse Anti-CaV1.2 Calcium Channel Monoclonal Antibody (SMC-300) at 1:1000.

<p>Immunohistochemistry analysis using Mouse Anti-CaV1.2 Calcium channel Monoclonal Antibody, Clone S57-47 (SMC-300). Tissue: Brain Tissue. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-CaV1.2 Calcium channel Monoclonal Antibody (SMC-300) at 1:10000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.</p>

Immunohistochemistry analysis using Mouse Anti-CaV1.2 Calcium channel Monoclonal Antibody, Clone S57-47 (SMC-300). Tissue: Brain Tissue. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-CaV1.2 Calcium channel Monoclonal Antibody (SMC-300) at 1:10000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Cav1.2 Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: SK-N-BE Cells (Human Neuroblastoma cells). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-Cav1.2 Monoclonal Antibody (SMC-300) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:200 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60 min at RT, 5 min at RT. Localization: Cell Membrane, Membrane, Cytoplasm, Nucleoplasm. Magnification: 60X. (A) Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain. (B) Anti-Cav1.2 Antibody. (C) Composite.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Cav1.2 Monoclonal Antibody, Clone S57-46 (SMC-300). Tissue: SK-N-BE Cells (Human Neuroblastoma cells). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-Cav1.2 Monoclonal Antibody (SMC-300) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:200 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60 min at RT, 5 min at RT. Localization: Cell Membrane, Membrane, Cytoplasm, Nucleoplasm. Magnification: 60X. (A) Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain. (B) Anti-Cav1.2 Antibody. (C) Composite.

Product Citations (2)

Other Citations

Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.

Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.

PubMed ID: N/A Reactivity Human Applications: Antibody Microarray

Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence.

Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22.

PubMed ID: N/A Reactivity Mouse Applications: Antibody Microarray

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

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

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amresco TAE缓冲液【1218】，amresco现货。Amresco公司来自美国，成立于 1976 年，为高质量生化试剂 / 试剂盒的生产商及供应商，产品服务于生物科研领域。用于体外诊断及医药中间体的美国 FDA 注册。amresco TAE缓冲液【1218】021-61806666 33779006品牌：AMRESCO数量：大量保存条件：4℃供应商：AMRESCO保质期：1年amresco TAE缓冲液【1218】CodeIt 查看更多>

平凡网吧_电话:05148716****

2021-07-16

pH计，是一种常用的仪器设备，主要用来精密测量液体介质的酸碱度值，配上相应的离子选择电极也可以测量离子电极电位MV值，pH计被广泛应用于环保、污水处理、科研、制药、发酵、化工、养殖、自来水等领域。该仪器也是食品厂、饮用水厂办QS、HACCP认证中的必备检验设备。... 查看更多>

德国Merck Dtube透析管说明书

2021-09-03

上海研生实业有限公司所提供的Citrate 柠檬酸盐缓冲液（0.01M pH6.0）质量可靠、规格齐全,上海研生实业有限公司不仅具有精湛的技术水平,更有良好的售后服务和优质的解决方案,欢迎您来电咨询此产品具体参数及价格等详细信息！查看更多>

智能制造基础核心国家标准《OPC统一架构》发布

2021-08-13

上海联硕生物科技有限公司在发布的TEN缓冲液(pH8.0)，10×TEN buffer 供应信息，浏览与TEN缓冲液(pH8.0)，10×TEN buffer 相关的产品或在搜索更多与TEN缓冲液(pH8.0)，10×TEN buffer 相关的内容。查看更多>

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商品咨询

缓冲溶液是什么,为啥可以维持溶液酸碱度稳定 123

知足3912017-10-02

【1】酸碱缓冲溶液：其中的一般酸碱缓冲溶液是指控制溶液酸碱度的溶液.他的作用是在此溶液中,加入少量的强酸或强碱,或者将其稍加稀释时,溶液的ph值基本上保持不变.【2】可以维持溶液酸碱度稳定的原因：主要原因是因为构成一般酸碱缓冲溶液的组分是“一元共轭酸碱”.例如“醋酸-醋酸钠”酸碱缓冲溶液.由PH的计算式：pH = p Ka +log[C酸/C碱] ,可见C酸/C碱的微小改变,其PH值,基本不变.

药典中,纯化水酸碱度能用PH计测量吗?《中国药典》2020版讨论区...123

rainbow09282021-07-25

药典上说取本品10ml，加甲基红指示液（变色范围ph4.2-6.3，红~黄）2滴不得显红色；加溴麝香草酚蓝（变色范围ph6.0-7.6，黄~蓝）5滴不得显蓝色。
是否可以理解为纯化水得PH范围为6.3-7.6？能否直接用pH计测量？谢谢!

酸碱滴定法中为什么要用缓冲溶液调节ph值123

格子控shy8032017-10-03

EDTA对金属离子的络合能力随酸度改变而不同，酸度越低，络合能力增强；酸度越高，络合能力越弱。可见控制溶液的pH在络合滴..

我是菜鸟,请教各位:强酸碱性样品可以用普通C18柱吗?谢谢了~ ...123

yandongliu2021-07-24

我的实验是考察药物不同PH值（水溶液，HCL和NaOH调PH值）下的稳定性情况等，PH从2-13。请问各位大侠：这种强酸性或强碱性水溶液样品可以直接进普通C18柱进行分析吗？
因为是考察不同PH对药物的影响，样品又不好改变其PH值，这种情况怎么办？希望有经验的高手指教。

我的流动相是甲醇-水（90：10）

谢谢赐教！

请进子版按格式发贴，自行修改，谢谢。

【求助】关于流动相添加剂分析技术论坛123

快乐药师2021-07-22

我们常用的色谱柱的PH耐受大概是PH（2-9），最近摸条件，但是pH计坏了，想调一下流动相的PH，苦于PH试纸的不精确性，特此求助：

常用流动相加酸碱后PH的总结，希望大家能够提供一点自己测过的结果，谢谢先

大家一起来讨论下哈~单抗分析:不同CEX方法测定同一个单抗,酸碱性...123

threetigers2015-03-06

如题：
两个CEX方法A和B测定同一单抗，结果碱性峰比例差不多，酸性峰比例相差约7%，相应主峰也差了7%左右。
具体来说，A方法酸性峰高，主峰低，碱性峰稍微低点；B方法酸性峰低，主峰高，碱性峰稍微高点；另外也做了CIEF，结果呢和A方法更接近。
仔细比较起来，AB两个方法的峰性和数量差不多，就不知道为什么会有这么大的差异。两个方法一个用的WCX柱-磷酸缓冲液，一个用SCX柱-MES缓冲液
大家帮我分析下：
1.两个方法哪个方法更准确，是以酸性峰高的为准还是什么？为什么？
2.这显著差异是由方法造成，具体原因是什么？柱子？
3.CIEF的结果和A方法更接近，是不是可以由此证明A方法更好或者CIEF的方法更好（因为CIEF更快更方便）？
欢迎讨论~
纠正下，A方法用的是Tosoh的柱子，B方法用的是SCX柱。TOSOH的柱子是7um的填料，10cm长。SCX是10um的填料。我本人TOSOH的阳离子柱子用的很少，这次信手用用，结果发现差异很大
那我现在就考虑，在以后方法开发过程中，除了通过流动相pH和组成、梯度、柱子选择来获得样品主峰和酸碱性的最大分离，还要关注各峰比例。因为之前比较方法好坏都只看分离度，尤其是主峰和邻近峰的分离度，获得最大分离度，自然可以做到主峰尽可能纯，但从未认真比较过各峰比例。这是一个大疏忽吧！
另外，CIEF和CEX方法原理还是有点差异的，所以分的是不同的异质体，原液放行两个方法肯定是都要做的。问题就是在早期细胞株筛选和工艺开发阶段，哪个方法才是又快又准。CIEF（iCE280)一般15分钟一个样，比CEX快多了。如果CIEF测得主峰要低于CEX结果，是不是真的完全可以取代CEX呢？CEX分离出的峰远比CIEF的多！
欢迎大家继续讨论~

一般的酸碱缓冲溶液有哪些类型,如何选择？123

2017-10-02

【酸碱缓冲溶液的类型及选择】酸碱缓冲溶液有三种类型：
1、弱酸和它的盐（如：HAc---NaAc）的水溶液组成；
2、弱碱和它的盐（如：NH3·H2O---NH4Cl）的水溶液组成；
3、多元弱酸的酸式盐及其对应的次级盐（如：NaH2PO4---Na2HPO4）的水溶液组成。
酸碱缓冲溶液的选型一般应根据具体情况进行选择。缓冲酸性可选用碱性缓冲液，缓冲酸性可采用碱性缓冲液。常用作缓冲溶液的酸类由弱酸及其共轭酸盐组合成的溶液具有缓冲作用。生化实验室常用的缓冲系主要有磷酸、柠檬酸、碳酸、醋酸、巴比妥酸、Tris（三羟甲基氨基甲烷）等系统，生化实验或研究工作中要慎重地选择缓冲体系，因为有时影响实验结果的因素并不是缓冲液的pH值，而是缓冲液中的某种离子。如硼酸盐、柠檬酸盐、磷酸盐和三羟甲基甲烷等缓冲剂都可能产生不需要的化学反应。
【酸碱缓冲溶液】由弱酸及其盐、弱碱及其盐组成的混合溶液，能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响，从而保持溶液的pH值相对稳定。这种溶液称为酸碱缓冲溶液。

为什么书上说:含有缓冲混合物的缓冲液实际上是含有同离子的弱酸...123

16凌凌凌凌凌2017-10-03

就是通过微弱的中和反应控制溶液的酸碱度。

磷酸钠缓冲液的配制方法 123

高展远瞩2014-10-12

看了GE公司的NiSephrose6FastFlow说明书，说bindingbuffer建议20mMsodiumphosphate,0.5MNaCl,pH7.4,elutingbuffer中是在bindingbuffer中加了高浓度的咪唑，我想问一下，这个怎么配制啊？那个sodiumphosphate是什么？磷酸钠溶液？貌似不具有缓冲性，磷酸钠缓冲液？有可能，但是这个咋配啊？有我能想到的三个配制方法，想求助一下;
1.直接用固体磷酸钠配制成50mM的磷酸钠溶液，再调pH到7.4；（我们试着用这个做了下，发现挂不上柱）
2.配置磷酸钠盐缓冲液：按NaH2PO4：Na2HPO4以19：81的摩尔比配制成pH7.4的缓冲液？（附一张百度出来的配方
）

3.如果是磷酸钠盐缓冲液，可以直接将50mM的NaH2PO4的水溶液用NaOH调成pH7.4吗？
再者，2和3这两个方法配制的磷酸钠盐缓冲液有什么区别？最终效果是一样的吗？如果不一样，有什么理论的知识支撑呢？个人感觉是分析化学中酸碱理论中的缓冲液那里的知识。求帮忙解答这些疑问。
另外，我还想问一下，pH对于Ni柱对His-tagged的蛋白的分离纯化影响大吗？是怎么影响的？谢谢大家了！

各类真空泵原理图解_二哈的世界你永远不懂!_真空泵原理123

2017-10-03

缓冲物质更重要。
由弱酸及其盐、弱碱及其盐组成的混合溶液，能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响，从而保持溶液的pH值相对稳定。这种溶液称为缓冲溶液。

凝胶电泳缓冲液的配制123

haina1682021-07-23

我按下面方法配制电泳缓冲液：
Tris-base15.1g
甘氨酸94g
SDS5g
H2O1000ml
听人说就这样配制，PH值就会在8.3左右，都不要怎么调PH的，但不知我这样配制后PH值在6.4左右？迷惑，配制了几次还是这样。本人初次做sds-page，请前辈们指教。
另外Tris是以前师兄留下的，几年了已经，SDS也是以前的，但是没有开封，密封的很好。甘氨酸是新买进口封装的。会不会试剂过期的原因？

求大侠帮助啊,怎样确定蛋白质的酸碱性啊??? 生物科学...123

空镜2021-07-20

请问什么软件可以测蛋白质的酸碱性