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即用型CDP-Star化学发光液 (0.4 mM, 含增强剂)
有货
为什么选择此级别
BioReagent,即用型,用于化学发光 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
即用型CDP-Star化学发光液（0.4mM,含增强剂）为即用型碱性磷酸酶(Alkaline Phosphatase, AP)化学发光底物，约10分钟内达到最大发光水平，辉光发射持续数小时，发射峰值波长位于460~470 nm。本品适用于基于膜的测定，可提高光输出和灵敏度，以出色的灵敏度、速度和易用性检测碱性磷酸酶和碱性磷酸酶标记分子。本化学发光底物为广泛使用的荧光底物甲基伞形酮磷酸酯（MUP）和比色底物对硝基苯磷酸盐（pNPP）提供了高度灵敏的替代物。低背景发光与高强度光输出相结合，能够以极高的灵敏度和信噪比检测碱性磷酸酶标记。
本品在基于膜的应用（例如Southern、Northern和Western印迹）中显示出高灵敏度。本品也可用于基于溶液的试验，如免疫检测、DNA探针试验、酶试验和报告基因试验。
产品应用
·Western Blot（AP标记二抗/探针）
·ELISA（AP标记检测抗体）
·核酸杂交（Southern/Northern Blot，AP标记探针）
·组织切片或细胞印迹（AP标记检测）
使用说明（以Western Blot为例，仅供参考）
1. 转膜后，用PBS或TBS简短漂洗印迹膜，并在至少10 mL封闭缓冲液中孵育30~60 min（或4℃过夜）。
注：PBS或TBS缓冲液均可用于硝酸纤维素膜或PVDF膜。对于尼龙膜，使用TBS的背景略高于PBS。
2. 用封闭缓冲液稀释一抗（5~10 mL），然后加入到印迹膜中。
3. 孵育30~60 min，然后洗涤至少2次，每次5 min。
对于硝酸纤维素和尼龙膜，使用洗涤缓冲液洗涤。
对于PVDF膜，使用封闭缓冲液洗涤。
注：所有洗涤步骤每次至少使用20 mL液体。
4. 用封闭缓冲液稀释二抗-AP结合物（参考抗体说明书推荐比例，通常1:1000~1:5000），然后加入到印迹膜中，孵育30~60 min。
5. 按步骤3的方法洗涤3次，每次5 min，最后用TBS（不含Tween-20）快速漂洗一次，去除表面活性剂（可降低背景）。
6. 加入CDP-Star发光液：
A. 将膜平铺于洁净保鲜膜或显影盒内，蛋白面朝上。
B. 用纸巾角吸干印迹膜上的多余液体（勿让膜干燥）。
C. 按每10cm2膜面积加入0.5~1 mL即用型CDP-Star发光液，确保完全覆盖。
D. 室温避光静置3~5 min，使底物充分反应。
7. 信号检测
CCD成像：直接将膜放入成像仪，曝光时间5s~5min（根据信号强弱调整）。
X光胶片：用吸水纸吸去多余发光液，保鲜膜包裹后压片，通常首次曝光0.5~2min。
注：使用PVDF或尼龙膜时的曝光时间通常短于硝酸纤维素膜。
注意事项
1. 本品开盖使用前请上下颠倒轻轻混匀。
2. 本品不宜在室温放置过久，以免影响检测效果，使用后请及时放回冰箱。
3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Ready‑to‑Use CDP‑Star Chemiluminescent Substrate (0.4 mM, with enhancer) is a ready‑to‑use chemiluminescent substrate for Alkaline Phosphatase (AP). Maximum light emission is reached within approximately 10 minutes, and the glow‑type signal can persist for several hours, with an emission peak wavelength of 460‑470 nm. Optimized for membrane‑based assays, this substrate delivers improved light output and sensitivity for the detection of alkaline phosphatase and AP‑labeled molecules with excellent sensitivity, speed and ease‑of‑use. It serves as a highly‑sensitive alternative to widely‑used fluorogenic substrate 4‑Methylumbelliferyl phosphate (MUP) and colorimetric substrate p‑Nitrophenyl phosphate (pNPP). The combination of low background luminescence and high‑intensity light output enables detection of AP‑labeled targets with ultra‑high sensitivity and signal‑to‑noise ratio.
High sensitivity is achieved in membrane‑based applications such as Southern‑blot, Northern‑blot and Western‑blot assays. This substrate is also suitable for solution‑based tests, including immunoassays, DNA probe assays, enzyme assays and reporter‑gene assays.
Applications
Western Blot (AP‑conjugated secondary antibodies / probes)
ELISA (AP‑labeled detection antibodies)
Nucleic acid hybridization (Southern / Northern Blot, AP‑labeled probes)
Tissue‑section or cell blot detection (AP‑based labeling)
Protocol (Western Blot as reference only)
1. After membrane transfer, briefly rinse the blot with PBS or TBS, and incubate in no less than 10 mL blocking buffer for 30‑60 min (or overnight at 4 °C).
Note: Both PBS and TBS are compatible with nitrocellulose and PVDF membranes. Slightly higher background may be observed on nylon membranes when TBS is used.
2. Dilute the primary antibody with blocking buffer (5‑10 mL total volume) and incubate the blot.
3. Incubate for 30‑60 min, then wash the blot at least twice for 5 min per wash.
Use wash buffer for nitrocellulose and nylon membranes.
Use blocking buffer for PVDF membranes.
Note: Use a minimum volume of 20 mL for each washing step.
4. Dilute the AP‑conjugated secondary antibody in blocking buffer (follow antibody supplier’s recommendations; typical dilution ratio: 1:1000‑1:5000), add to the blot and incubate for 30‑60 min.
5. Wash 3 times for 5 min each as described in Step 3, followed by a quick final rinse in TBS (without Tween‑20) to remove surfactant and reduce background signal.
6. Add CDP‑Star substrate solution
A. Place the blot flat on clean plastic wrap or inside an incubation cassette, protein‑side facing upward.
B. Gently blot excess liquid from the edge of the membrane with tissue paper. Do not allow the membrane to dry out.
C. Apply 0.5‑1 mL ready‑to‑use CDP‑Star substrate per 10 cm² of membrane surface to ensure full coverage.
D. Incubate the blot at room temperature in the dark for 3‑5 min to allow sufficient substrate reaction.
7. Signal detection
CCD imager: Place the blot directly into the imaging system; adjust exposure time from 5 s to 5 min according to signal intensity.
X‑ray film: Remove excess substrate solution, wrap the blot in plastic wrap and expose to X‑ray film. An initial exposure of 0.5‑2 min is recommended.
Note: PVDF or nylon membranes generally require shorter exposure times than nitrocellulose membranes.
Precautions
1. Invert and mix gently up‑and‑down before opening and using the product.
2. Do not leave the substrate at room temperature for extended periods, which may impair detection performance. Return the reagent to refrigerator storage promptly after use.
3. For personal safety, perform all operations while wearing a lab coat and disposable gloves.
规格
规格或纯度
BioReagent, 即用型, 用于化学发光, 0.4mM
稳定性与储存
Store at 2-8°C long term (12 months).
英文名称
Ready‑to‑Use CDP‑Star Chemiluminescent Substrate (0.4 mM, with enhancer)
储存条件
2-8°C储存
运输条件
低温运输
Emission(nm)
470nm
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
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技术文档和文章
溶液计算器
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