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StressMarq/Anti-CD74 Antibody [1B8]/SMC-267D-A565/100-µg

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StressMarq/Anti-CD74 Antibody [1B8]/SMC-267D-A565/100-µg

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SMC-267D-A565

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Overview:

Product Name CD74 Antibody

Description

Mouse Anti-Human CD74 Monoclonal IgG2a

Species Reactivity Human

Applications WB, ICC/IF

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

Host Species Mouse

Immunogen Species Human

Immunogen Human CD74 invariant chain synthetic peptide.

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	Protein G Purified
Clonality	Monoclonal
Clone Number	1B8
Isotype	IgG2a
Specificity	Detects ~33 kDa protein.
Cite This Product	StressMarq Biosciences Cat# SMC-267, RRID: AB_2699952
Certificate of Analysis	A 1:1000 dilution of SMC-267 was sufficient for detection of CD74 in 15 µg of B-Cell Lymphoma cell line Raji by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary Antibody.

Biological Description

Alternative Names	DHLAG Antibody, HLA DR gamma Antibody, HLADG Antibody, p33 Antibody, p35 Antibody, Protein 41 Antibody, CD 74 Antibody, CD74 Antibody, CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) Antibody, CD74 antigen Antibody, CD74 molecule Antibody, CD74 molecule, major histocompatibility complex, class II invariant chain Antibody, CLIP Antibody, DHLAG Antibody, Gamma chain of class II antigens Antibody, HG2A_HUMAN Antibody, HLA class II histocompatibility antigen gamma chain Antibody, HLA DR antigens associated invariant chain Antibody, HLA DR gamma Antibody, HLA-DR antigens-associated invariant chain Antibody, HLA-DR-gamma Antibody, HLADG Antibody, HLADR antigens associated invariant chain Antibody Ia antigen associated invariant chain Antibody, Ia antigen-associated invariant chain Antibody, Ia associated invariant chain Antibody, Ia gamma Antibody, Ii Antibody, Invariant polypeptide of major histocompatibility complex class II antigen associated Antibody, la-gamma Antibody, Major histocompatibility complex class II invariant chain Antibody, MHC HLA DR gamma chain Antibody, MHC HLA-DR gamma chain Antibody, p33 Antibody, p35 Antibody, Protein 41 Antibody
Research Areas	Adaptive Immunity, B Cell CD Markers, B Cells, Immunology
Cellular Localization	Cell membrane, Endoplasmic Reticulum, Endoplasmic reticulum membrane, Endosome, Golgi apparatus, Lysosome
Accession Number	NP_001020329.1
Gene ID	972
Swiss Prot	P04233
Scientific Background	CD74 is a non-polymorphic type II integral membrane protein. It has a short N-terminal cytoplasmic tail of 28 amino acids, followed by a single 24-aa transmembrane region and an approximately 150-aa lumenal domain (1). The CD74 chain is thought to function mainly as an MHC class II chaperone, which promotes ER exit of MHC class II molecules, directs them to endocytic compartments, prevents peptide binding in the ER, and contributes to peptide editing in the MHC class II compartment. Class II MHC and Ii expression was believed to be restricted to classical antigen-presenting cells (APC); however, during inflammation, other cell types, including mucosal epithelial cells, have also been reported to express class II MHC molecules (2). Experiments that investigate cell-surface CD74 are complicated by the fact that CD74 remains on the cell surface for a very short time. The surface half-life of CD74 was calculated to be fewer than 10 minutes (3). CD74 however has also recently been shown to have a role as an accessory-signaling molecule because of its high-affinity binding to the pro-inflammatory cytokine, macrophage migration-inhibitory factor (MIF) (3). The restricted expression of CD74 by normal tissues and its very rapid internalization make CD74 an attractive therapeutic target for both cancer and immunologic diseases (4).
References	1. Becker-Hermann, S., Arie, G., Medvedovsky H, Kerem A, and Shachar I. (2005) Mol Bio Cell. 16(11):5061-9. 2. Barrera CA., et al (2005) J Histochem Cytochem 53 (12): 1481-9. 3. Starlets D., et al. (2006) Blood. 107 (12): 4807-4816. 4. Burton JD., et al. (2004). Clin Cancer Res. 10(19): 6606-11. 5. Denzin L.K., Hammond, C. and Cresswell, P. (1996) J. Exp. Med. 184: 2153-2165. 6. Denzin L.K., Robbins N.F., Carboy-Newcomb C. and Cresswell P. (1994) Immunity 1: 595-606.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Tissue: HeLa Cells (Human Cervical Cancer). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) 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, Endoplasmic Reticulum, Golgi apparatus. Magnification: 60X. (A) DAPI (blue) nuclear stain (B) Phalloidin Texas Red F-Actin stain (C) CD74 Antibody (D) Composite.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Tissue: HeLa Cells (Human Cervical Cancer). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) 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, Endoplasmic Reticulum, Golgi apparatus. Magnification: 60X. (A) DAPI (blue) nuclear stain (B) Phalloidin Texas Red F-Actin stain (C) CD74 Antibody (D) Composite.

<p>Western Blot analysis of Human Lymphoblastoid cell line (Raji) showing detection of 33-35 kDa CD74 protein using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Lane 1: Molecular Weight Ladder (MW). Lane 2: Raji cell lysate. Load: 15 µg. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:1000 for 60 min at RT. Color Development: ECL solution for 5 min in RT. Predicted/Observed Size: 33-35 kDa.</p>

Western Blot analysis of Human Lymphoblastoid cell line (Raji) showing detection of 33-35 kDa CD74 protein using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Lane 1: Molecular Weight Ladder (MW). Lane 2: Raji cell lysate. Load: 15 µg. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:1000 for 60 min at RT. Color Development: ECL solution for 5 min in RT. Predicted/Observed Size: 33-35 kDa.

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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

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

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2021-07-28

上海笃玛生物科技有限公司在发布的TESCA缓冲液(pH7.4,无菌)供应信息，浏览与TESCA缓冲液(pH7.4,无菌)相关的产品或在搜索更多与TESCA缓冲液(pH7.4,无菌)相关的内容。查看更多>

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商品咨询

如何选择缓冲溶液?_123

sweetwater2021-07-21

做试验的时候用到很多种缓冲液，Tris,triethanolamine,HEPES,还有磷酸缓冲液，不知道这些缓冲液之间有没有大的区别，选择缓冲液的依据是什么？

如果只是为了缓冲酸碱度，是不是可以用一种缓冲液来代替所有其他的？这样做试验就方便多了。谢谢指教

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.

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

【求助】Whatman的DEAE52到底用不用酸碱预处理?怎么说法不一...123

langfeng2021-07-26

Whatman的网站上说DE52（货号4057-200）是预膨胀的（Preswollen），有的中文网站上说DE52是即用型。
这就是说不用酸碱预处理吗？
Whatman的网站上没有DE52最大耐受压力，请问又经验的战友应该是多少？

Whatman的网站上：
DE32DryMicrogranularDEAECellulose
SimilarperformancecharacteristicsafterprecyclingasDE52.

DE52PreswollenMicrogranularDEAECellulose
ProbablythemostwidelyusedDEAEcelluloseintheworld;usedforbiopolymerswithlowtohighnegativecharges;exhibitsexcellentresolutionwithgoodflowrates.

附件是一本图书(MethodsinMolecularMedicine,)的章节,上面说:
WhatmanDEAE52comesalreadypreswollenandonlyneedstobetransferred
totherunningbuffer50mMTE8.

lAntibodiesUsingIonExchangeChromatography.pdf(87.06k)

缓冲溶液就是能抵抗外来酸碱影响,保持pH值绝对不变的溶液。 () ...123

linaqb134662017-10-02

缓冲溶液为什么能够低抗外来酸碱对ph的改变

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。

PH值在多少为碱性水?123

wuzhixuan20032005-06-13

25倍电转缓冲液的ph＝8.3真的不能调吗？听师兄说一次配好液后它的ph值就必须是8.3，不能再进行加酸或加碱调节，可我配了一天也没达到这个要求，用的是18g甘氨酸和3.64g的trisbase加dd水到100ml，总是ph值到8.8左右，在配过程中可以加热吗，不然太难溶解了，

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？

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!

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~

培养基高压蒸气灭菌后pH值的变化及其对细菌生长的影响123

mars7472005-08-25

:)
我在做一细菌不同酸碱度生长状况时,发现这些奇怪现象:pH=3的培养基灭菌(TSB液体培养基)灭菌后pH上升到到9.2!而原来pH=9.0的降到8.7(基本没多少变化),请问各位大侠,这是什么原因?

一般做不同酸碱度生长实验时,该如何才能防止pH在湿热灭菌后基本不变化?

【求助】蛋白质酸碱性对离子交换色谱填料的选择蛋白质和糖学...123

ldx74226632009-08-05

求助：我目前需用离子交换法进行蛋白质的分离和纯化，但是我不知道要纯化的蛋白质的等电点，怎么样选择柱料及洗脱缓冲液呢？如果必须知道，有没有什么简便的方法测定蛋白质的等电点呢？谢谢