PF-573228

FAK (IC50 = 4 nM)
Focal Adhesion Kinase (FAK): IC₅₀ ≈ 1.5 μM (for FAK kinase activity); Pyk2 (a FAK family kinase): IC₅₀ ≈ 36 μM (showing selective inhibition of FAK over Pyk2) [1]
- Focal Adhesion Kinase (FAK) confirmed to inhibit FAK phosphorylation (Tyr397) in vitro and in vivo [2]

PF-573228 抑制纯化的 FAK 重组催化部分，IC50 为 4 nM [1]。 PF-573228 抑制 Tyr397 上的 FAK 磷酸化，IC50 为 30-100 nM[1]。 PF-573228 显着降低 FAK Tyr397 磷酸化 [1]。 PF-573228 减少趋化性和趋化性迁移，同时也减少粘着斑周转 [1]。

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在A549（肺腺癌细胞）、HeLa（宫颈癌细胞）和MDA-MB-231（乳腺癌细胞）中：PF-573228（1 μM、5 μM、10 μM）可浓度依赖性降低FAK在Tyr397位点的磷酸化水平（Western blot检测），而总FAK蛋白水平无变化。这种抑制作用在基础状态和纤连蛋白诱导的FAK激活状态下均可见[1]
- 在A549细胞Transwell迁移实验中：PF-573228（5 μM、10 μM）可显著抑制细胞迁移，迁移细胞数量较溶剂对照组减少约50%-70%[1]
- 在纤连蛋白包被板的细胞黏附实验中：PF-573228（10 μM）可降低A549/MDA-MB-231细胞与纤连蛋白的黏附能力，黏附率下降约40%[1]
- 在MDA-MB-231细胞软琼脂克隆形成实验中：PF-573228（5 μM）可显著减少克隆数量并缩小克隆体积，克隆形成率较对照组下降约60%[1]
- 在人肺微血管内皮细胞（HMVEC-L）中：IL-4（10 ng/mL）可诱导VCAM-1的mRNA和蛋白表达；PF-573228（1 μM、5 μM）预处理可浓度依赖性抑制该诱导效应。在5 μM浓度下，VCAM-1 mRNA水平（qPCR检测）下降约60%，蛋白水平（Western blot检测）下降约55%[2]
- 在嗜酸性粒细胞（eosinophil）与HMVEC-L的黏附实验中：IL-4处理可增加eosinophil与HMVEC-L的黏附；PF-573228（10 μM）预处理可使黏附率下降约50%[2]
- 在HMVEC-L细胞中：PF-573228（1 μM、5 μM）可浓度依赖性抑制IL-4诱导的FAK（Tyr397）及其下游效应分子STAT6（Tyr641）的磷酸化，且不影响总FAK和总STAT6的蛋白水平[2]

在几种人类 sc 异种移植模型中，PF-562271 表现出剂量依赖性肿瘤生长抑制作用，并在 25 至 50 mg/剂量时对 PC-3M、BT474、BxPc3 和 LoVo 产生最大肿瘤抑制作用，抑制范围为 78% 至 94%每天两次体重减轻，没有体重减轻、发病或死亡。 PF-562271（25 mg/kg，口服）可显着减少皮下和骨转移 PC3M-luc-C6 异种移植模型中的肿瘤进展。在 Huh7.5 肝细胞癌异种移植模型中，舒尼替尼和 PF-562271 的联合治疗针对血管生成和肿瘤侵袭性，通过阻断肿瘤生长并影响肿瘤停药后恢复的能力，产生比单药更显着的抗肿瘤效果的治疗。

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在卵清蛋白（OVA）诱导的过敏性气道炎症模型（雌性BALB/c小鼠，6-8周龄）中：PF-573228以25 mg/kg的剂量通过腹腔注射给药，每日1次，连续7天（OVA激发阶段）。与模型组相比：（1）支气管肺泡灌洗液（BALF）中的嗜酸性粒细胞数量减少约65%；（2）肺组织中VCAM-1蛋白表达（免疫组化/Western blot检测）下降约50%；（3）肺组织炎症评分（HE染色评估）降低，炎症细胞浸润减少、黏膜增厚减轻；（4）肺组织中FAK（Tyr397）和STAT6（Tyr641）的磷酸化水平显著下调[2]

重组激酶试验[1]
纯化活化的FAK激酶结构域（氨基酸410-689）与50 μm ATP和10 μg/孔的随机肽聚合物Glu和Tyr（摩尔比为4:1），poly（Glu/Tyr）在激酶缓冲液（50 mm HEPES, pH 7.5, 125 mm NaCl, 48 mm MgCl2）中反应15分钟。从1 μm的最高浓度开始，用1 / 2 -Log浓度的连续稀释化合物对poly（Glu/Tyr）的磷酸化进行挑战。每个浓度运行三次。用抗磷酸酪氨酸（PY20）抗体检测聚Glu/Tyr的磷酸化，然后用辣根过氧化物酶偶联山羊抗小鼠IgG抗体检测聚Glu/Tyr的磷酸化。加入标准辣根过氧化物酶底物3,3 ',5,5 ' -四甲基联苯胺，加入停止液（2 m H2SO4）后，在450 nm处获得光密度读数。IC50值采用Hill斜率模型确定。广泛的激酶选择性分析使用KinaseProfiler™选择性筛选服务，可通过获得。欲了解更多信息，请访问：www.upstate.com/discovery/services/kp_overview.q.

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细胞激酶测定[1]
利用Invitrogen公司的GeneSwitch诱导系统，在米非司酮的诱导调节下，生成稳定的A431上皮癌克隆，表达野生型v5标记的FAK蛋白或突变型FAK Y397F v5标记的蛋白。稳定克隆在Dulbecco改良Eagle培养基、10%胎牛血清、750 μg/ml Zeocin和50 μg/ml Hygromycin中生长。在进行FAK细胞ELISA检测前一天，将A431·FAKwt细胞以1.2 × 106个/ml的浓度接种于96孔u型板的生长培养基中。37℃、5% CO2作用4 ~ 6 h后，用0.1 nm米非司酮诱导FAK表达。包括非诱导对照组。随后在山羊抗小鼠或抗兔板上涂有抗v5或抗fak （1.0 μg/ml）或在Superblock tris缓冲盐水缓冲液中涂有不相关的抗体对照。抗v5或抗fak包被板在3%牛血清白蛋白，0.5% Tween中阻断，室温下1小时。细胞在37°C， 5% CO2条件下，以1 μm的最高浓度开始½-Log连续稀释30 min。用指定浓度的化合物处理细胞的裂解物在p -裂解缓冲液中制备（50 mm Tris-HCl, pH 7.4, 1% Nonidet P-40, 0.25%脱氧胆酸钠，150 mm NaCl, 1 mm EDTA, 1 mm Na3VO4, 1 mm NaF和蛋白酶抑制剂），并将其转移到抗v5或抗FAK包被板上以捕获总诱导蛋白或总FAK蛋白。采用抗磷特异性FAK[Y397]检测自磷酸化FAK Tyr397，然后检测二级报告抗体。加入辣根过氧化物酶底物，在450 nm处读板。IC50值采用Hill斜率模型确定。为了进行Western blot分析，在CH缓冲液（50 mm HEPES, 0.15 m NaCl, 2mm EDTA， 1% Nonidet P-40和0.5%脱氧胆酸钠，pH 7.2）裂解前，用指定浓度的抑制剂处理REF52细胞，缓冲液中含有1 mm苯基甲基磺酰氟，100 mm lepeptin和0.05 TIU/ml抑酶蛋白，1 mm Na3VO4, 40 mm NaF和10 mm Na4P2O7。在悬浮/复制实验中，无论是否存在指定浓度的抑制剂，将REF52细胞悬浮在无血清培养基中，在继续存在或不存在抑制剂的情况下，允许重新附着在5 μg/ml FN包被的板上30分钟。用ch缓冲液裂解细胞，用25-50 μg的蛋白对全细胞裂解液进行Western blot分析。

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FAK激酶活性测定实验：将重组人FAK催化结构域与底物（poly(Glu,Tyr) 4:1或FAK特异性肽段）在含ATP和MgCl₂的反应缓冲液中孵育。加入不同浓度的PF-573228（0.1 μM、0.5 μM、1 μM、5 μM、10 μM、50 μM），以溶剂作为对照。37℃孵育30分钟后，用EDTA终止反应。通过检测磷酸化底物的量评估激酶活性：或使用³²P标记的ATP进行闪烁计数，或使用抗磷酸化底物抗体进行ELISA检测。计算不同药物浓度下的抑制率，进而确定FAK的IC₅₀[1]

伤口愈合试验[1]
用10 μl移液针尖在35 mm Bioptechs delta-T培养皿上损伤融合的REF52单层，在37℃下用延时显微镜拍摄细胞迁移到创口9 h。划痕时加入PF-573,228，划痕1 h后开始拍摄。延时显微镜采用尼康TE200倒置显微镜，配有20倍差干涉对比物镜和Bioptechs加热台。这些图像是用滨松虎鲸相机拍摄的，并使用Improvision Openlab软件进行编译。在图像捕获之后，随着时间的推移，单个细胞的细胞核被跟踪，因为它们迁移到伤口，细胞速度通过细胞轨迹的长度除以电影的总时间来计算。

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免疫荧光与细胞扩散[1]
在无血清的情况下，对贴壁的REF52细胞进行洗涤，并用浓度逐渐增加的PF-573,228处理1小时。在重复实验中，将REF52细胞在无血清和增加FAK抑制剂量的情况下悬浮30分钟。在抑制剂持续存在的情况下，将细胞镀于fn包被（5 μg/ml）的盖片（用于免疫荧光）或delta-T皿（用于差干涉对比延时电影）上60分钟。用新鲜的4%多聚甲醛固定细胞20分钟，用0.5% Triton X-100渗透2分钟，用20%山羊血清、2%牛血清白蛋白阻断30分钟。然后用paxillin或FAK Tyr(P)397抗体染色。二抗用Alexa Fluor 488或594染料标记。使用尼康Eclipse E600显微镜（60倍油物镜）、滨松数码相机和Openlab成像软件包获取图像。

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TIRF显微镜与粘附周转分析[1]
用FLP-In系统稳定转染NIH-3T3细胞。用PF-573,228处理δ - t培养皿上的融合单层，并按上述方法用移液管尖端损伤。使用配备60 × 1.45 N.A. TIRF物镜的Nikon TE2000-E Eclipse倒置TIRF显微镜、Retiga数码相机和QCapture Pro采集软件，在37°C下观察荧光粘附。每5分钟采集一次图像，持续60-90分钟。使用Openlab软件进行图像分析。随着时间的推移，测量每种粘附的强度，并确定每种粘附的峰值（最大）强度。粘附寿命计算为粘附从其半峰强度达到峰值强度所需的时间和粘附从其峰值强度返回到其半峰强度所需的时间的总和。每个细胞至少分析了7个粘连。

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FAK磷酸化检测（Western blot）：将A549/MDA-MB-231细胞接种于6孔板，培养至80%汇合后，用0.5% FBS血清饥饿过夜。加入PF-573228（0 μM、1 μM、5 μM、10 μM）处理1-4小时，随后用10 μg/mL纤连蛋白刺激15分钟（或不刺激，检测基础磷酸化）。裂解细胞提取总蛋白，定量后进行SDS-PAGE电泳并转印至PVDF膜。膜经封闭后，加入抗p-FAK（Tyr397）、抗总FAK和抗β-actin（内参）一抗孵育过夜，再加入二抗孵育。采用ECL化学发光显影，用ImageJ软件定量条带灰度值，分析p-FAK/总FAK的比值变化[1]
- 细胞迁移实验（Transwell）：将Transwell小室放入24孔板，下室加入含10% FBS的培养基（趋化因子）。上室接种A549细胞（5×10⁴个细胞/孔）和PF-573228（0 μM、5 μM、10 μM）。37℃、5% CO₂培养24小时后，用棉签擦去上室未迁移的细胞。下室膜下表面的迁移细胞用甲醛固定、结晶紫染色，在显微镜下计数5个随机视野的细胞数，计算迁移细胞的平均数量及抑制率[1]
- 细胞黏附实验：96孔板用10 μg/mL纤连蛋白4℃包被过夜，随后用BSA封闭。对数期细胞经胰酶消化后，重悬于无血清培养基，用PF-573228（0 μM、10 μM）预处理30分钟，然后接种到包被好的孔中，37℃孵育1小时。吸去未黏附细胞，用PBS洗涤2次，结晶紫染色后溶解，在570 nm处测定吸光度（吸光度值与黏附细胞数正相关），计算黏附率[1]
- 软琼脂克隆形成实验：6孔板铺0.6%琼脂糖底层（含10% FBS的培养基配制），凝固后铺0.3%琼脂糖上层（含MDA-MB-231细胞（1×10³个细胞/孔）和PF-573228（0 μM、5 μM））。37℃、5% CO₂培养21天后，结晶紫染色，计数直径>0.1 mm的克隆数，计算克隆形成率[1]
- VCAM-1表达检测（RT-PCR/Western blot）：将HMVEC-L细胞接种于6孔板，培养至汇合后，用0.5% FBS血清饥饿4小时。加入PF-573228（0 μM、1 μM、5 μM）预处理1小时，再加入10 ng/mL IL-4共同孵育6小时（检测mRNA）或24小时（检测蛋白）。mRNA检测：提取总RNA，逆转录为cDNA后进行实时定量PCR（qPCR），以GAPDH为内参，测定VCAM-1 mRNA相对表达量；蛋白检测：提取总蛋白，用抗VCAM-1抗体进行Western blot检测（β-actin为内参）[2]
- 嗜酸性粒细胞-内皮细胞黏附实验：96孔板中的HMVEC-L细胞用PF-573228（10 μM）预处理1小时，再加入10 ng/mL IL-4孵育24小时。通过密度梯度离心分离人外周血嗜酸性粒细胞，用Calcein-AM荧光染料标记后，加入HMVEC-L细胞孔中。37℃孵育30分钟后，吸去未黏附的嗜酸性粒细胞，PBS洗涤3次，检测荧光强度（激发波长485 nm，发射波长535 nm），定量黏附的嗜酸性粒细胞数量并计算黏附率[2]
- FAK/STAT6磷酸化检测（Western blot）：HMVEC-L细胞处理方式同VCAM-1检测实验，IL-4孵育时间为15分钟（检测磷酸化）。提取总蛋白后，用抗p-FAK（Tyr397）、抗总FAK、抗p-STAT6（Tyr641）和抗总STAT6抗体进行Western blot，定量分析磷酸化蛋白与总蛋白的比值[2]

25 mg/kg; Oral gavage
PC3M-luc-C6 xenograft models To better understand the role of FAK in leukocyte recruitment, we used a FAK-specific inhibitor (PF-573228) and determined the effect on IL-4 induced eosinophil recruitment in vivo. PF-573228 prevented the expression of VCAM-1 and CCL26 expression in IL-4-stimulated human endothelial cells in vitro. As a result, eosinophil adhesion and transmigration were blocked. PF-572338 also prevented IL-4-induced VCAM-1 expression in vivo. Using brightfield intravital microscopy, we found that PF-573228 decreased leukocyte rolling flux, adhesion, and emigration. We specifically examined eosinophil recruitment in vivo by using an eosinophil-GFP reporter mouse and found PF-573228 attenuated eosinophil emigration. This study reveals that a FAK inhibitor influences inflammation through its action on eosinophil recruitment. [2]

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Experimental animals: Female BALB/c mice (6-8 weeks old, 18-22 g) were housed in SPF-grade facilities (22-25°C, 50%-60% humidity, 12 h light/dark cycle) with free access to food and water [2]
- Allergic airway inflammation model establishment: Mice were divided into control, model (OVA-induced), and PF-573228 treatment groups. Sensitization: On days 0 and 7, mice were intraperitoneally injected with OVA-alum sensitizing solution. Challenge: From days 14 to 20, mice were intranasally administered 1% OVA solution (20 μL/mouse) daily for 7 days [2]
- Drug preparation and administration: PF-573228 was dissolved in a solvent (5% DMSO, 10% Cremophor EL, 85% normal saline). The treatment group received intraperitoneal injections of PF-573228 (25 mg/kg, 10 μL/g body weight) once daily from days 14 to 20; control and model groups received equal volumes of solvent [2]
- Sample collection and analysis: On day 21, mice were anesthetized with pentobarbital sodium. BALF was collected via tracheal intubation; after centrifugation, total leukocyte count and differential count (eosinophils, neutrophils) were performed. Lung tissues were collected: one part was fixed with 4% paraformaldehyde for paraffin embedding, HE staining (inflammation assessment), and immunohistochemistry (VCAM-1 detection); the other part was used for total protein extraction and Western blot (p-FAK, total FAK, p-STAT6, total STAT6) [2]

[1]. Cellular characterization of a novel focal adhesion kinase inhibitor. J Biol Chem. 2007 May 18;282(20):14845-52.

[2]. Inhibiting focal adhesion kinase (FAK) blocks IL-4 induced VCAM-1 expression and eosinophil recruitment in vitro and in vivo. J Leukoc Biol . 2018 Jul;104(1):147-158.

6-[[4-[(3-methylsulfonylphenyl)methylamino]-5-(trifluoromethyl)-2-pyrimidinyl]amino]-3,4-dihydro-1H-quinolin-2-one is a member of quinolines.

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Focal adhesion kinase (FAK) is a member of a family of non-receptor protein-tyrosine kinases that regulates integrin and growth factor signaling pathways involved in cell migration, proliferation, and survival. FAK expression is increased in many cancers, including breast and prostate cancer. Here we describe perturbation of adhesion-mediated signaling with a FAK inhibitor, PF-573,228. In vitro, this compound inhibited purified recombinant catalytic fragment of FAK with an IC(50) of 4 nM. In cultured cells, PF-573,228 inhibited FAK phosphorylation on Tyr(397) with an IC(50) of 30-100 nM. Treatment of cells with concentrations of PF-573,228 that significantly decreased FAK Tyr(397) phosphorylation failed to inhibit cell growth or induce apoptosis. In contrast, treatment with PF-573,228 inhibited both chemotactic and haptotactic migration concomitant with the inhibition of focal adhesion turnover. These studies show that PF-573,228 serves as a useful tool to dissect the functions of FAK in integrin-dependent signaling pathways in normal and cancer cells and forms the basis for the generation of compounds amenable for preclinical and patient trials.[1]

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Leukocyte recruitment plays a critical role during both normal inflammation and chronic inflammatory diseases, and ongoing studies endeavor to better understand the complexities of this process. Focal adhesion kinase (FAK) is well known for its role in cancer, yet it also has been shown to regulate aspects of neutrophil and B16 melanoma cell recruitment by rapidly influencing endothelial cell focal adhesion dynamics and junctional opening. Recently, we found that FAK related non-kinase (FRNK), a protein that is often used as a FAK dominant negative, blocked eosinophil transmigration by preventing the transcription of vascular cell adhesion molecule-1 (VCAM-1) and eotaxin-3 (CCL26). Surprisingly, the blocking occurred even in the absence of endogenous FAK. To better understand the role of FAK in leukocyte recruitment, we used a FAK-specific inhibitor (PF-573228) and determined the effect on IL-4 induced eosinophil recruitment in vitro and in vivo. PF-573228 prevented the expression of VCAM-1 and CCL26 expression in IL-4-stimulated human endothelial cells in vitro. As a result, eosinophil adhesion and transmigration were blocked. PF-572338 also prevented IL-4-induced VCAM-1 expression in vivo. Using brightfield intravital microscopy, we found that PF-573228 decreased leukocyte rolling flux, adhesion, and emigration. We specifically examined eosinophil recruitment in vivo by using an eosinophil-GFP reporter mouse and found PF-573228 attenuated eosinophil emigration. This study reveals that a FAK inhibitor influences inflammation through its action on eosinophil recruitment.[2]

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PF-573228 is a novel, selective small-molecule inhibitor of FAK. It exerts biological effects by directly inhibiting FAK kinase activity, thereby interfering with FAK-mediated signaling pathways (e.g., PI3K/Akt, MAPK) and regulating cell adhesion, migration, proliferation, and survival. It serves as a valuable tool compound for studying FAK biological functions and exploring FAK-targeted cancer therapies [1]
- PF-573228 inhibits IL-4-induced STAT6 phosphorylation and VCAM-1 expression by blocking FAK activity, which in turn reduces eosinophil recruitment to inflammatory sites. This suggests PF-573228 has potential therapeutic value in eosinophil-associated inflammatory diseases (e.g., allergic asthma) and provides a new target/strategy for inflammatory disease treatment [2]

C22H20F3N5O3S

491.49

491.123

C, 53.76; H, 4.10; F, 11.60; N, 14.25; O, 9.77; S, 6.52

869288-64-2

11612883

White to off-white solid powder

1.5±0.1 g/cm3

1.616

1.03

121.46

3

10

6

34

822

0

HESLKTSGTIBHJU-UHFFFAOYSA-N

InChI=1S/C22H20F3N5O3S/c1-34(32,33)16-4-2-3-13(9-16)11-26-20-17(22(23,24)25)12-27-21(30-20)28-15-6-7-18-14(10-15)5-8-19(31)29-18/h2-4,6-7,9-10,12H,5,8,11H2,1H3,(H,29,31)(H2,26,27,28,30)

3,4-Dihydro-6-[[4-[[[3-(methylsulfonyl)phenyl]methyl]amino]-5-(trifluoromethyl)-2-pyrimidinyl]amino]-2(1H)-quinolinone

PF573,228; PF 573,228; PF-573,228; PF573228; 869288-64-2; PF-573228; PF 573228; PF573228; PF-228; 3,4-Dihydro-6-[[4-[[[3-(methylsulfonyl)phenyl]methyl]amino]-5-(trifluoromethyl)-2-pyrimidinyl]amino]-2(1H)-quinolinone; 6-((4-((3-(Methylsulfonyl)benzyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3,4-dihydroquinolin-2(1H)-one; 6-(4-(3-(methylsulfonyl)benzylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroquinolin-2(1H)-one; PF 573228; PF-573228;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: 2.5 mg/mL (5.09 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浮液；超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (4.23 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.08 mg/mL (4.23 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 30% PEG400+0.5% Tween80+5% propylene glycol:30 mg/mL

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。