ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
CCL3-L1; LD78; SCYA3L; SCYA3L1; MIP1AP; MIP1-Alpha-P; Small Inducible Cytokine A3-Like 1; Macrophage Inflammatory Protein 1-Alpha-P; Tonsillar Lymphocyte LD78 Beta
- Product No.SEA026Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- All
- Human
- Mouse
- Rat
- Cavia
- Rabbit
- Simian
- Caprine
- Ovine
- Equine
- Bovine
- Porcine
- Gallus
- Canine
- Others
- Multi-species
- Pan-species
- Test MethodDouble-antibody Sandwich
- Assay Length3h
- Detection Range0.156-10ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.055ng/mL.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 441 For more details, please contact local distributors!US$ 630 For more details, please contact local distributors!US$ 2835 For more details, please contact local distributors!US$ 5355 For more details, please contact local distributors!US$ 44100 For more details, please contact local distributors!
Specificity of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
This assay has high sensitivity and excellent specificity for detection of Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1).No significant cross-reactivity or interference between Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) and analogues was observed.
Recovery of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
Matrices listed below were spiked with certain level of recombinant Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) and the recovery rates were calculated by comparing the measured value to the expected amount of Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 79-91 | 81 |
EDTA plasma(n=5) | 85-94 | 89 |
heparin plasma(n=5) | 93-102 | 97 |
Precision of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>10%>Inter-Assay: CV<12%>12%>
Linearity of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 83-102% | 78-101% | 98-105% | 95-105% |
EDTA plasma(n=5) | 92-99% | 86-101% | 85-103% | 89-98% |
heparin plasma(n=5) | 85-94% | 83-93% | 95-102% | 87-102% |
Stability of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Assay procedure summary of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
1. Prepare all reagents, samples and standards;2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;4. Aspirate and wash 3 times;5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;6. Aspirate and wash 5 times;7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle of the ELISA Kit for Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1)
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Chemokine C-C-Motif Ligand 3 Like Protein 1 (CCL3L1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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Catalog No. | Organism species: Homo sapiens (Human) | Applications (RESEARCH USE ONLY!) |
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RAW细胞,6孔板,RIPA提取蛋白(提前加好PMAF),每孔100uLRIPA,冰上用tip倒过来刮细胞,放置15min,12000gX15min离心,取上清。做WB时,沸水煮5min,胶冻样物质还是存在。
操作过程中发现离心后还是有胶冻样物质,和上清混在一起无法分开,严重的时候基本上全是胶冻样物质。
这个胶冻样物质应该是DNA,园友提供的方法为用超声碎裂器,但是我们实验室没有。请问还有别的方法可以使用么?
如要膜蛋白保持原来的结构应采用:非离子型去垢剂.
分子式:C14H22O(C2H4O)n
用途:Triton X-100是一种比较温和的去垢剂(表面活性剂或称界面活性剂),常作为添加剂使蛋白保持稳定,尤其是膜蛋白.
性质:
1.Triton X-100对细菌等微生物没有杀伤作用.
2.Triton X-100在紫外波段下有光吸收(lambda max = 275 nm and 283 nm in methanol),因此,当缓冲液中存在Triton X-100时不能通过测定280nm光吸收来进行蛋白定量.
3.Triton X-100常与CHAPS等zwitterionic 去垢剂一起使用来纯化膜蛋白,使膜蛋白保持其天然构象.
4.Triton X-100可以提高真核细胞细胞膜的通透性,因此在Immunostaining时常使用含 0.1%-0.5%Triton X-100的PBS或TBS.
5.Triton X-100非常稳定,密封避光 +2 to +8℃条件下可以长期保存.
6.Triton X-100是透明,略显粘稠,稍微发黄的液体.
7.Triton X-100的密度为1.07 g/ml,pH6.0-8.0(5%的溶液).
8.Triton X-100可以高压灭菌.
9.用Triton X-100来裂解细胞时,0.1% Triton X-100就足够了,不过,达到0.5%时也没问题.
10.蛋白酶K在含1% Triton X-100的溶液中依然保持活性.
11.可以使用Amberlite hydrophobic XAD resins and Rezorian A161 cartridges 来去除Triton X-100 .
去垢剂(又称表面活性剂)是一类即具有亲水基又具有疏水基的物质,一般具有乳化、分散、和增溶作用,可分阴离子、阳离子和中性去垢剂等多种类型,中性去垢剂在蛋白提取中应用的较多.
中性去垢剂又称非离子表面活性剂,对蛋白质的变性作用影响较少,宜于蛋白质或酶提取之用.一般市售中性去垢剂有聚乙二醇类,如PEG200;多元醇类表面活性剂,如山梨醇、司盘类和吐温类;聚氧乙烯脂肪醇醚,如苄泽类、平平加类;聚氧乙烯烷基苯酚醚,如Igepal CO、乳化剂OP、Triton、Pluronic(用作消泡剂、润湿剂、增溶剂)、泡敌.中性去垢剂作用后可通过Sephadex LH-50柱除去;也可直接上DEAE-Sephadex柱层析分离目的蛋白,不必先除去去垢剂.
如常用于缓冲的Tris、HEPES、MPOS等等,去垢剂如NP40,Tween-20,TritonX100,CHAPS,脱氧胆酸钠,SDS等等,
大家能说说这些试剂的使用区别吗(最好讲讲某些情况下使用一些会优于另一些的例子)
有时候某些裂解液含10%左右甘油,又是为何?
平时我们用习惯了一些配方,可能有时候不太清楚为何有些是可替代的,有些是不可替代的,借此机会想听听大家的见解?
去垢剂作为添加剂生长蛋白质晶体是一项很有用的技术。非离子和两性离子结晶剂在膜蛋白结晶中已经得到很好应用,目前已经变成常规的实验手段。去垢剂在一些可溶蛋白中也起到了一定的作用,提高了晶体的质量和结果的可重复性 ,促进了单晶的生长, 但具体的影响机制尚不明确。其可能原因是去垢剂的引入使结晶液滴中的蛋白分子更具有亲水性,使溶液中同质蛋白质分子增加。向左转|向右转