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引用資料與參考文獻 (9)
Invitrogen™
Alexa Fluor™ 430 NHS Ester (Succinimidyl Ester)
Alexa Fluor™ 430 is a bright, green-fluorescent dye. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor™深入閱讀
| 產品號碼 | Quantity |
|---|---|
| A10169 | 5 mg |
產品號碼 A10169
價格 (HKD)
3,994.00
Each
Quantity:
5 mg
價格 (HKD)
3,994.00
Each
Alexa Fluor™ 430 is a bright, green-fluorescent dye. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor™ 430 dye is water soluble and pH-insensitive from pH 4 to pH 10. In addition to reactive dye formulations, we offer Alexa Fluor™ 430 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates optimized for cellular labeling and detection.
The NHS ester (or succinimidyl ester) of Alexa Fluor™ 430 is the most popular tool for conjugating this dye to a protein or antibody. NHS esters can be used to label to the primary amines (R-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting Alexa Fluor™ conjugate will exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores.
Detailed information about this AlexaFluor™ NHS ester:
Fluorophore label: Alexa Fluor™ 430 dye
Reactive group: NHS ester
Reactivity: Primary amines on proteins and ligands, amine-modified oligonucleotides
Ex/Em of the conjugate: 430/545 nm
Extinction coefficient: 15,000 cm-1M-1
Molecular weight: 701.8
Typical Conjugation Reaction
You can conjugate amine-reactive reagents with virtually any protein or peptide (the provided protocol is optimized for IgG antibodies). You can scale the reaction for any amount of protein, but the concentration of the protein should be at least 2 mg/mL for optimal results. We recommend trying three different degrees of labeling, using three different molar ratios of the reactive reagent to protein.
The Alexa Fluor™ NHS ester is typically dissolved in high-quality anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO) (D12345), and the reaction is carried out in 0.1–0.2 M sodium bicarbonate buffer, pH 8.3, at room temperature for 1 hour. Because the pKa of the terminal amine is lower than that of the lysine epsilon-amino group, you may achieve more selective labeling of the amine terminus using a buffer closer to neutral pH.
Conjugate Purification
Labeled antibodies are typically separated from free Alexa Fluor™ dye using a gel filtration column, such as Sephadex™ G-25, BioGel™ P-30, or equivalent. For much larger or smaller proteins, select a gel filtration media with an appropriate molecular weight cut-off or purify by dialysis. We offer several purification kits optimized for different quantities of antibody conjugate:
Antibody Conjugate Purification Kit for 0.5-1 mg (A33086)
Antibody Conjugate Purification Kit for 20-50 μg (A33087)
Antibody Conjugate Purification kit for 50-100 μg (A33088)
Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See our Antibody Labeling kits or use our Labeling Chemistry Selection Tool for other choices. To learn more about our labeling kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in The Molecular Probes™ Handbook.
We’ll Make a Custom Conjugate for You
If you can’t find what you’re looking for in our online catalog, we’ll prepare a custom antibody or protein conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 13485:2000 certified.
The NHS ester (or succinimidyl ester) of Alexa Fluor™ 430 is the most popular tool for conjugating this dye to a protein or antibody. NHS esters can be used to label to the primary amines (R-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting Alexa Fluor™ conjugate will exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores.
Detailed information about this AlexaFluor™ NHS ester:
Fluorophore label: Alexa Fluor™ 430 dye
Reactive group: NHS ester
Reactivity: Primary amines on proteins and ligands, amine-modified oligonucleotides
Ex/Em of the conjugate: 430/545 nm
Extinction coefficient: 15,000 cm-1M-1
Molecular weight: 701.8
Typical Conjugation Reaction
You can conjugate amine-reactive reagents with virtually any protein or peptide (the provided protocol is optimized for IgG antibodies). You can scale the reaction for any amount of protein, but the concentration of the protein should be at least 2 mg/mL for optimal results. We recommend trying three different degrees of labeling, using three different molar ratios of the reactive reagent to protein.
The Alexa Fluor™ NHS ester is typically dissolved in high-quality anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO) (D12345), and the reaction is carried out in 0.1–0.2 M sodium bicarbonate buffer, pH 8.3, at room temperature for 1 hour. Because the pKa of the terminal amine is lower than that of the lysine epsilon-amino group, you may achieve more selective labeling of the amine terminus using a buffer closer to neutral pH.
Conjugate Purification
Labeled antibodies are typically separated from free Alexa Fluor™ dye using a gel filtration column, such as Sephadex™ G-25, BioGel™ P-30, or equivalent. For much larger or smaller proteins, select a gel filtration media with an appropriate molecular weight cut-off or purify by dialysis. We offer several purification kits optimized for different quantities of antibody conjugate:
Antibody Conjugate Purification Kit for 0.5-1 mg (A33086)
Antibody Conjugate Purification Kit for 20-50 μg (A33087)
Antibody Conjugate Purification kit for 50-100 μg (A33088)
Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See our Antibody Labeling kits or use our Labeling Chemistry Selection Tool for other choices. To learn more about our labeling kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in The Molecular Probes™ Handbook.
We’ll Make a Custom Conjugate for You
If you can’t find what you’re looking for in our online catalog, we’ll prepare a custom antibody or protein conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 13485:2000 certified.
For Research Use Only. Not for use in diagnostic procedures.
規格
Chemical ReactivityAmine
Label or DyeAlexa Fluor™ 430
Quantity5 mg
Reactive MoietyActive Ester, Succinimidyl Ester
Shipping ConditionRoom Temperature
Label TypeAlexa Fluor
Product LineAlexa Fluor
Unit SizeEach
內容物與存放
Store in freezer (-5 to -30°C) and protect from light.
常見問答集 (常見問題)
I am labeling a protein with Alexa Fluor 488 SDP ester. The manual recommends using a sodium bicarbonate buffer at pH 8.3. Can I use a different buffer instead?
I am not going to use all of my Alexa Fluor succinimidyl ester reactive dye. Can I just make it up in DMSO and store aliquots at -20 degrees C?
引用資料與參考文獻 (9)
引用資料與參考文獻
Abstract
Small heat shock protein Hsp16.3 modulates its chaperone activity by adjusting the rate of oligomeric dissociation.
Journal:Biochem Biophys Res Commun
PubMed ID:14521926
'Small heat shock proteins usually exist as oligomers and appear to undergo dynamic dissociation/reassociation, with oligomeric dissociation being a prerequisite for their chaperone activities. However, contradictory cases were also reported that chaperone activities could be enhanced with no change or even increase in oligomeric sizes. Using Hsp16.3 as a model
Detection and quantification of biotinylated proteins using the Storm 840 Optical Scanner.
Journal:J Nutr Biochem
PubMed ID:12770643
'The use of the avidin-biotin interaction is becoming an increasingly common method for the detection of proteins. The use of fluorescence detection with avidin-biotin systems has the potential to greatly increase both the sensitivity and linearity of this type of analysis. In this report, three fluorescent systems were tested for
In vivo blockade of the programmed cell death-1 pathway using soluble recombinant PD-1-Fc enhances CD4+ and CD8+ T cell responses but has limited clinical benefit.
Journal:
PubMed ID:24227774
'The programmed cell death-1 (PD-1)/programmed cell death ligand-1 pathway has been shown to limit cell-mediated effector functions during chronic viral infections impeding clearance of pathogens. As a strategy to reverse this exhaustion and increase T cell polyfunctionality, PD-1 ligands were blocked in vivo using a recombinant macaque PD-1 fused to
Platelets support a protective immune response to LCMV by preventing splenic necrosis.
Journal:Blood
PubMed ID:22566603
Severe arenaviral infections in humans are characterized by clinical findings common to other viral hemorrhagic fevers (VHFs), including thrombocytopenia, leukopenia, skin and internal organ hemorrhages, high viral replication, splenic necrosis, and death. Host responses, rather than direct damage by the arenaviral replication, account for most of the observed pathology, but
Plasmacytoid dendritic cells are recruited to the colorectum and contribute to immune activation during pathogenic SIV infection in rhesus macaques.
Journal:Blood
PubMed ID:21693759
In SIV/HIV infection, the gastrointestinal tissue dominates as an important site because of the impact of massive mucosal CD4 depletion and immune activation-induced tissue pathology. Unlike AIDS-susceptible rhesus macaques, natural hosts do not progress to AIDS and resolve immune activation earlier. Here, we examine the role of dendritic cells (DCs)