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StressMarq/Anti-KDEL Antibody/SPC-109B-A390/200-µg

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StressMarq

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SPC-109B-A390

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Overview:

Product Name KDEL Antibody

Description

Rabbit Anti-KDEL Polyclonal

Species Reactivity Human, Mouse, Rat

Applications WB, ICC/IF, IHC

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

Host Species Rabbit

Immunogen KDEL containing peptide immunogen

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 pH7.2, 50% glycerol, 0.09% sodium azide
Storage Temperature	-20ºC
Shipping Temperature	Blue Ice or 4ºC
Purification	Protein A purified
Clonality	Polyclonal
Specificity	Detects KDEL proteins, GRP94, Grp78, PDI and calreticulin. It may also see ERp57 and ERp29.
Cite This Product	StressMarq Biosciences Cat# SPC-109, RRID: AB_10805031
Certificate of Analysis	A 1:1000 dilution of SPC-109 was sufficient for detection of KDEL-containing proteins in 20 µg of HeLa cell lysate by ECL immunoblot analysis using goat anti-mouse IgG as the secondary.

Biological Description

Alternative Names	Lys Asp Glu Leu Antibody
Research Areas	Cell Signaling, Chaperone Proteins, Organelle Markers, Protein Trafficking, Tags and Cell Markers
Cellular Localization	Endoplasmic Reticulum
Scientific Background	The endoplasmic reticulum is part of a protein sorting pathway, or in essence, the transportation system of the eukaryotic cell. The majority of endoplasmic reticulum resident proteins are retained in the endoplasmic reticulum through a retention motif. This motif is composed of four amino acids at the C-terminal end of the protein sequence. The most common retention sequence is KDEL (lys-asp-glu-leu). Grp78 and Grp94 and PDI all share the C-terminal KDEL sequence. The presence of carboxy-terminal KDEL appears to be necessary for ER retention and appears to be sufficient to reduce the secretion of proteins from the ER.
References	1. Ozawa K., et al. (2008) Mol Pharmacol. 74:1610. 2. Austin R.C., et al. (2003) J Biol Chem. 278: 17438.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-KDEL Polyclonal Antibody (SPC-109). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-KDEL Polyclonal Antibody (SPC-109) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-KDEL Antibody. (C) Composite. Heat Shocked at 42°C for 30 min.</p>

Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-KDEL Polyclonal Antibody (SPC-109). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-KDEL Polyclonal Antibody (SPC-109) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-KDEL Antibody. (C) Composite. Heat Shocked at 42°C for 30 min.

<p>Western blot analysis of Human Cell line lysates showing detection of KDEL protein using Rabbit Anti-KDEL Polyclonal Antibody (SPC-109). Primary Antibody: Rabbit Anti-KDEL Polyclonal Antibody (SPC-109) at 1:1000, 1:500.</p>

Western blot analysis of Human Cell line lysates showing detection of KDEL protein using Rabbit Anti-KDEL Polyclonal Antibody (SPC-109). Primary Antibody: Rabbit Anti-KDEL Polyclonal Antibody (SPC-109) at 1:1000, 1:500.

Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-KDEL Polyclonal Antibody (SPC-109). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-KDEL Polyclonal Antibody (SPC-109) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-KDEL Antibody. (C) Composite. Heat Shocked at 42°C for 30 min.

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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

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

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【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

短期内输库存血,病人容易发生的酸碱平衡紊乱是() 123

fdsez2015-08-06

相关疾病：高钾血症碱中毒代谢性酸中毒输入库存血，钾离子大量释放入血，会导致高钾血症，同时容易伴发代谢性碱中毒，查了相关资料解释说：大量输...

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！

酸碱加入到缓冲液做的excel的散点图怎么做123

怪小米米2017-10-03

首先你需要有源数据，
有了源数据之后把源数据按照大小排列，
选中源数据区域-->ALT+A1-->选中图标区右键-->更改图表类型-->散点图

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。

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。

【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

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？

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?

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

一种缓冲溶液是一个共轭酸碱的混合物,那么为什么邻苯二甲酸氢钾、四硼 ...123

TWDAFRA01242017-10-03

缓冲溶液是含共轭酸碱对的，其中共轭酸与共轭碱都必须是弱酸或弱碱，这样才能组成缓冲液。因为邻苯二甲酸是弱酸，氨是弱碱，并且它们的同浓度的电离度相似，所以组成的缓冲溶液缓冲能力强。

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!