- Description
- Additional
Description
Details
Description:Mouse monoclonal antibody (mAb) to human cluster of differentiation 11 alpha chain (CD11α) or mAb to human lymphocyte function associated factor I (LFA-1) alpha chain.
Purification:Protein G affinity purified
Product Type:Primary antibody
Target Protein:Human CD11α
Immunogen:Human cytolytic T lymphocytes
Fusion Myeloma:Sp2/0-Ag14
Specificity:This antibody is reactive to human CD11α.
Species Reactivity:Human, others not tested.
Host / Isotype:Mouse, IgG1 Kappa
Formulation:Lyophilized from a solution in 0.01M PBS, pH 7.0
Reconstitution:Double distillated water is recommended to adjust the final concentration to 1.00mg/mL.
Storage: Store at -20oC
Research Area:Immunology
Background:
CD11α comprises the alpha-chain of lymphocyte function associated factor I (LFA-1). LFA-1 is an adhesion molecule expressed on the surface of T cell, B cell, macrophage, monocytes and neutrophils. LFA-1 plays a role in lymphocyte recruitment to site of infection. LFA-1 initiates a weak link between lymphocytes and antigen presentation cells (APC) by binding to intercellular adhesion molecule 1 (ICAM-1) on APC.
Application:
Fluorescence flow cytometry:Above (see image) is the histogram of Jurkat cells stained with 10mg/ml mouse anti-human CD11α monoclonal antibody. Black line histogram represents Jurkat cells stained with isotype control, mouse anti-TB 38Kd antigen monoclonal antibody (MO-I40021D, B12F8).
Immuno-histochemistry
References:
If research is published using this product, please inform Anogen in order to cite the reference on this datasheet. Anogen will provide one unit of product in the same category as gratitude.
Additional
Additional Information
| Product Specificity | mAb anti-Human CD11α, A11 |
|---|---|
| Application | IF, IHC |
| Size | 0.1 mg |
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生物偶联即将两种分子结合在一起而形成一个杂合的偶联物(hybridconjugate),这些偶联结合物(如标记抗体)在世界各大医院和科研机构被广泛使用。一抗标记试剂系列产品极大程度的简化了结合标记流程,是目前世界上操作最简便的抗体标记试剂盒。
·快速:手工操作时间仅30秒·目前世界上操作最简便的抗体标记试剂盒·无需脱盐、透析等步骤·抗体回收率达100%·可提供数十种标记物·可用于WB,IHC,ELISA,免疫荧光、流式细胞术以及荧光共振能量转移检测等·经济实惠,规格多样·可标记ug-mg级抗体量试剂盒组成:
1)1或3(以规格而定)玻管Lightening-Link?Mix混合物。
2)1管LL-X(X为标记物,如PE或HRP等)修饰试剂。
3)1管LL-X萃灭试剂。
操作步骤:
1)在每10ul抗体样品中加入1ul的LL-Modifier修饰试剂,轻轻混匀。
2)打开Ligtening-Link混合物管盖,吸取步骤1已混好的抗体样品至管内冻干混合物中,用吸液器轻轻吸打1-2次,使粉末完全溶解。
3)盖上玻璃瓶盖,室温(20-25°C)避光孵育3小时,或过夜。*
4)孵育3小时后,按每10ul抗体的量加入1ulLL-淬灭试剂,静置30分钟后即可使用。
*不同的标记物,操作步骤略有不同。(操作步骤供参考,以英文原版说明书为准。)
标记结合物的保存:
对于任何新结合物,推荐保持于4°C条件下。如需长期保存,可添加保护剂。
缓冲液及添加剂兼容性:
A.与无胺基缓冲液兼容,包括MES,MOPS,HEPES(浓度10-50mM,pH范围6.5-8.5),LL-Modifier修饰剂提供结合反应的最佳条件。
B.常见的盐(如氯化钠)、螯合剂(EDTA)、糖对偶联反应无影响。
C.叠氮钠(0.02%-0.1%)对结合反应有轻微影响,叠氮钠不可用于HRP标记,其为后者的抑制剂。
D.甘油(upto50%)对结合效率无影响。
E.若无胺基的缓冲液浓度相对较高,pH值在6.6-8.5范围外,则需要加入更多的LL-midifier修饰剂(/10ul抗体),以此来调整pH值。最佳的pH范围为7.3-7.6左右。
F.避免使用含亲核剂的缓冲液,避免还原剂如DTT和2-巯基乙醇,乙醇胺等。
G.浓度达到或稍低于20mM的Tris对结合效率有轻微的影响。
Westernblottingdatacomparingconjugatedandunconjugatedantibodies
货号中文品名英文品名规格AZK0001AColorlockALS磷检测试剂盒PiColorlockALS625/1560assaysAZK0001BColorlockALS磷检测试剂盒PiColorlockALS2500/6250assaysAZK0002AColorlockGold磷检测试剂盒PiColorlockGold625/1560assaysAZK0002BColorlockGold磷检测试剂盒PiColorlockGold2500/6250assaysAZK0003APi结合树脂PiBind5gAZK0004AATP琼脂糖Low(1-2umol/ml)ATPagaroseLow(1-2umol/ml)2mlAZK0004BATP琼脂糖Low(1-2umol/ml)ATPagaroseLow(1-2umol/ml)5mlAZK0005AATP琼脂糖High(>6umol/ml)GTPagaroseHigh(>6umol/ml)2mlAZK0006AATP琼脂糖High(1-2umol/ml)GTPagaroseHigh(>6umol/ml)5mlAZK0007AATP琼脂糖High(8-12umol/ml)ATPagaroseHigh(8-12umol/ml)2mlAZK0008AATP琼脂糖High(8-12umol/ml)ATPagaroseHigh(8-12umol/ml)5mlAZK0009A控制树脂ControlResin2mlAZK0009B控制树脂ControlResin5mlAZK0010AATPase酶检测试剂盒ATPaseassaysystem192assaysAZK0010BATPase酶检测试剂盒ATPaseassaysystem480assaysAZK0010CATPase酶检测试剂盒ATPaseassaysystem5x480assaysAZK0011AGTPase酶检测试剂盒GTPaseassaysystem192assaysAZK0011BGTPase酶检测试剂盒GTPaseassaysystem480assaysAZK0011CGTPase酶检测试剂盒GTPaseassaysystem5x480assaysAZK0013AAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit3x10μgAPAZK0013BAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit1x100μgAPAZK0013CAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit3x100μgAPAZK0013DAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit1mgAPAZK0014AGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit3x10μgAZK0014BGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit3x100μgAZK0014CGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit1x1mgAZK0015AR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)3x10μgPEAZK0015BR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)1x100μgPEAZK0015CR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)3x100μgPEAZK0015DR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)1mgPEAZK0016AB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)1x100μgBPEAZK0016BB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)3x100μgBPEAZK0016CB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)1mgBPEAZK0017AAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)3x10μgAPCAZK0017BAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)1x100μgAPCAZK0017CAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)3x100μgAPCAZK0017DAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)1mgAPCAZK0018APerCP标记试剂盒Lightning-LinkPerCP3x10ugAZK0018BPerCP标记试剂盒Lightning-LinkPerCP1x100ugAZK0018CPerCP标记试剂盒Lightning-LinkPerCP3x100ugAZK0018DPerCP标记试剂盒Lightning-LinkPerCP1x1mgAZK0019A荧光素标记试剂盒Lightning-LinkFluorescein3reactions(eachupto20ug)AZK0019B荧光素标记试剂盒Lightning-LinkFluorescein3reactions(eachupto200ug)AZK0019C荧光素标记试剂盒Lightning-LinkFluorescein1reaction(upto2mg)AZK0020A罗丹明标记试剂盒Lightning-LinkRhodamine3reactions(eachupto20ug)AZK0020B罗丹明标记试剂盒Lightning-LinkRhodamine3reactions(eachupto200ug)AZK0020C罗丹明标记试剂盒Lightning-LinkRhodamine1reaction(upto2mg)AZK0021AAMCA标记试剂盒Lightning-LinkAMCA3reactions(eachupto20ug)AZK0021BAMCA标记试剂盒Lightning-LinkAMCA3reactions(eachupto200ug)AZK0021CAMCA标记试剂盒Lightning-LinkAMCA1reaction(upto2mg)AZK0022ATexasRed标记试剂盒Lightning-LinkTexasRed3reactions(eachupto20ug)AZK0022BTexasRed标记试剂盒Lightning-LinkTexasRed3reactions(eachupto200ug)AZK0022CTexasRed标记试剂盒Lightning-LinkTexasRed1reaction(upto1mgscale)AZK0023ACy3染料标记试剂盒Lightning-LinkCy33reactions(eachupto20ug)AZK0023BCy3染料标记试剂盒Lightning-LinkCy33reactions(eachupto200ug)AZK0023CCy3染料标记试剂盒Lightning-LinkCy31reaction(upto2mg)AZK0024ACy5染料标记试剂盒Lightning-LinkCy53reactions(eachupto20ug)AZK0024BCy5染料标记试剂盒Lightning-LinkCy53reactions(eachupto200ug)AZK0024CCy5染料标记试剂盒Lightning-LinkCy51reaction(upto2mg)AZK0025ACy5.5染料标记试剂盒Lightning-LinkCy5.53reactions(eachupto200ug)AZK0025BCy5.5染料标记试剂盒Lightning-LinkCy5.51reaction(upto2mg)AZK0026ACy7染料标记试剂盒Lightning-LinkCy73reactions(eachupto20ug)AZK0026BCy7染料标记试剂盒Lightning-LinkCy73reactions(eachupto200ug)AZK0026CCy7染料标记试剂盒Lightning-LinkCy71reaction(upto2mg)AZK0027ACy3.5染料标记试剂盒Lightning-LinkCy3.53reactions(eachupto200ug)AZK0027BCy3.5染料标记试剂盒Lightning-LinkCy3.51reaction(upto2mg)AZK0028AAtto390荧光染料标记试剂盒Lightning-Linkatto3903reactions(eachupto20ug)AZK0028BAtto390荧光染料标记试剂盒Lightning-Linkatto3903reactions(eachupto200ug)AZK0028CAtto390荧光染料标记试剂盒Lightning-Linkatto3901reaction(upto2mg)AZK0029AAtto425荧光染料标记试剂盒Lightning-Linkatto4253reactions(eachupto20ug)AZK0029BAtto425荧光染料标记试剂盒Lightning-Linkatto4253reactions(eachupto200ug)AZK0029CAtto425荧光染料标记试剂盒Lightning-Linkatto4251reaction(upto2mg)AZK0030AAtto465荧光染料标记试剂盒Lightning-Linkatto4653reactions(eachupto20ug)AZK0030BAtto465荧光染料标记试剂盒Lightning-Linkatto4653reactions(eachupto200ug)AZK0030CAtto465荧光染料标记试剂盒Lightning-Linkatto4651reaction(upto2mg)AZK0031AAtto488荧光染料标记试剂盒Lightning-Linkatto4883reactions(eachupto20ug)AZK0031BAtto488荧光染料标记试剂盒Lightning-Linkatto4883reactions(eachupto200ug)AZK0031CAtto488荧光染料标记试剂盒Lightning-Linkatto4881reaction(upto2mg)AZK0032AAtto532荧光染料标记试剂盒Lightning-Linkatto5323reactions(eachupto20ug)AZK0032BAtto532荧光染料标记试剂盒Lightning-Linkatto5323reactions(eachupto200ug)AZK0032CAtto532荧光染料标记试剂盒Lightning-Linkatto5321reaction(upto2mg)AZK0033AAtto565荧光染料标记试剂盒Lightning-Linkatto5653reactions(eachupto20ug)AZK0033BAtto565荧光染料标记试剂盒Lightning-Linkatto5653reactions(eachupto200ug)AZK0033CAtto565荧光染料标记试剂盒Lightning-Linkatto5651reaction(upto2mg)AZK0034AAtto590荧光染料标记试剂盒Lightning-Linkatto5903reactions(eachupto20ug)AZK0034BAtto590荧光染料标记试剂盒Lightning-Linkatto5903reactions(eachupto200ug)AZK0034CAtto590荧光染料标记试剂盒Lightning-Linkatto5901reaction(upto2mg)AZK0035AAtto594荧光染料标记试剂盒Lightning-Linkatto5943reactions(eachupto20ug)AZK0035BAtto594荧光染料标记试剂盒Lightning-Linkatto5943reactions(eachupto200ug)AZK0035CAtto594荧光染料标记试剂盒Lightning-Linkatto5941reaction(upto2mg)AZK0036AAtto633荧光染料标记试剂盒Lightning-Linkatto6333reactions(eachupto20ug)AZK0036BAtto633荧光染料标记试剂盒Lightning-Linkatto6333reactions(eachupto200ug)AZK0036CAtto633荧光染料标记试剂盒Lightning-Linkatto6331reaction(upto2mg)AZK0037AAtto637荧光染料标记试剂盒Lightning-Linkatto6373reactions(eachupto200ug)AZK0037BAtto637荧光染料标记试剂盒Lightning-Linkatto6371reaction(upto2)mgAZK0038AAtto655荧光染料标记试剂盒Lightning-Linkatto6553reactions(eachupto20ug)AZK0038BAtto655荧光染料标记试剂盒Lightning-Linkatto6553reactions(eachupto200ug)AZK0038CAtto655荧光染料标记试剂盒Lightning-Linkatto6551reaction(upto2mg)AZK0039AAtto680荧光染料标记试剂盒Lightning-Linkatto6803reactions(eachupto20ug)AZK0039BAtto680荧光染料标记试剂盒Lightning-Linkatto6803reactions(eachupto200ug)AZK0039CAtto680荧光染料标记试剂盒Lightning-Linkatto6801reaction(upto2mg)AZK0040AAtto700荧光染料标记试剂盒Lightning-Linkatto7003reactions(eachupto20ug)AZK0040BAtto700荧光染料标记试剂盒Lightning-Linkatto7003reactions(eachupto200ug)AZK0040CAtto700荧光染料标记试剂盒Lightning-Linkatto7001reaction(upto2mg)AZK0041AFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H3reactions(eachupto20ug)AZK0041BFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H3reactions(eachupto200ug)AZK0041CFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H1reaction(upto2mg)AZK0042AFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5943reactions(eachupto20ug)AZK0042BFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5943reactions(eachupto200ug)AZK0042CFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5941reaction(upto2mg)AZK0043AFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H3reactions(eachupto20ug)AZK0043BFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H3reactions(eachupto200ug)AZK0043CFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H1reaction(upto2mg)AZK0044AFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6823reactions(eachupto20ug)AZK0044BFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6823reactions(eachupto200ug)AZK0044CFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6821reaction(upto2mg)AZK0045AFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7523reactions(eachupto20ug)AZK0045BFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7523reactions(eachupto200ug)AZK0045CFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7521reaction(upto2mg)AZK0046APE/Cy5串联染料Lightning-LinkPE/Cy53x10ugAZK0046BPE/Cy5串联染料Lightning-LinkPE/Cy51x100ugAZK0046CPE/Cy5串联染料Lightning-LinkPE/Cy53x100ugAZK0046DPE/Cy5串联染料Lightning-LinkPE/Cy51x1mgAZK0047APE/Cy5.5串联染料Lightning-LinkPE/Cy5.53x10ugAZK0047BPE/Cy5.5串联染料Lightning-LinkPE/Cy5.51x100ugAZK0047CPE/Cy5.5串联染料Lightning-LinkPE/Cy5.53x100ugAZK0047DPE/Cy5.5串联染料Lightning-LinkPE/Cy5.51x1mgAZK0048APE/Cy7串联染料Lightning-LinkPE/Cy73x10ugAZK0048BPE/Cy7串联染料Lightning-LinkPE/Cy71x100ugAZK0048CPE/Cy7串联染料Lightning-LinkPE/Cy73x100ugAZK0048DPE/Cy7串联染料Lightning-LinkPE/Cy71x1mgAZK0049APE/Cy5.5串联染料Lightning-LinkPerCP/Cy5.53x10ugAZK0049BPE/Cy5.5串联染料Lightning-LinkPerCP/Cy5.51x100ugAZK0050APE/TexasRed串联染料Lightning-LinkPE/TexasRed3x10ugAZK0050BPE/TexasRed串联染料Lightning-LinkPE/TexasRed1x100ugAZK0050CPE/TexasRed串联染料Lightning-LinkPE/TexasRed3x100ugAZK0050DPE/TexasRed串联染料Lightning-LinkPE/TexasRed1x1mgAZK0051APE/Atto594串联染料Lightning-Link?PE/Atto5943x10ugAZK0051BPE/Atto594串联染料Lightning-Link?PE/Atto5941x100ugAZK0051CPE/Atto594串联染料Lightning-Link?PE/Atto5943x100ugAZK0051DPE/Atto594串联染料Lightning-Link?PE/Atto5941x1mgAZK0052APerCP/Cy5.5串联染料Lightning-LinkPerCP/Cy5.53x100ugAZK0052BPerCP/Cy5.5串联染料Lightning-LinkPerCP/Cy5.51x1mgAZK0052CPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.53x10ugAZK0052DPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.51x100ugAZK0052EPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.53x100ugAZK0052FPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.51x1mgAZK0053ALightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy73x10ugAZK0053BLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy71x100ugAZK0053CLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy73x100ugAZK0053DLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy71x1mgAZK0054A生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA3reactions(upto20μgAb)AZK0054B生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA3reactions(upto200μgAb)AZK0054C生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA1reaction(upto2mgAb)AZK0055A生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB3reactions(upto20μgAb)AZK0055B生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB3reactions(upto200μgAb)AZK0055C生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB1reaction(upto2mgAb)AZK0056A链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit3x10ugStreptavidinAZK0056B链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit3x100ugStreptavidinAZK0056C链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit1x1mgStreptavidinAZK0057A亲和素标记试剂盒Lightning-LinkAvidinconjugationkit3x10ugAvidinAZK0057B亲和素标记试剂盒Lightning-LinkAvidinconjugationkit3x100ugAvidinAZK0057C亲和素标记试剂盒Lightning-LinkAvidinconjugationkit1x1mgAvidinAZK0058A快速荧光素标记Lightning-Link?RapidFluorescein3xAblabelings(each10-20ugscale)AZK0058B快速荧光素标记Lightning-Link?RapidFluorescein3xAblabelings(each100-200ugscale)AZK0058C快速荧光素标记Lightning-Link?RapidFluorescein1xAblabeling(1scale)AZK0059A快速Cy3标记Lightning-Link?RapidCy33xAblabelings(each10-20ugscale)AZK0059B快速Cy3标记Lightning-Link?RapidCy33xAblabelings(each100-200ugscale)AZK0059C快速Cy3标记Lightning-Link?RapidCy31xAblabeling(1scale)AZK0060A快速Cy5标记Lightning-Link?RapidCy53xAblabelings(each10-20ugscale)AZK0060B快速Cy5标记Lightning-Link?RapidCy53xAblabelings(each100-200ugscale)AZK0060C快速Cy5标记Lightning-Link?RapidCy51xAblabeling(1scale)AZK0061A快速Atto488标记Lightning-Link?RapidAtto4883xAblabelings(each10-20ugscale)AZK0061B快速Atto488标记Lightning-Link?RapidAtto4883xAblabelings(each100-200ugscale)AZK0061C快速Atto488标记Lightning-Link?RapidAtto4881xAblabeling(1scale)AZK0062A快速Atto633标记Lightning-Link?RapidAtto6333xAblabelings(each10-20ugscale)AZK0062B快速Atto633标记Lightning-Link?RapidAtto6333xAblabelings(each100-200ugscale)AZK0062C快速Atto633标记Lightning-Link?RapidAtto6331xAblabeling(1scale)AZK0063A快速Atto565标记Lightning-Link?RapidAtto5653xAblabelings(each10-20ugscale)AZK0063B快速Atto565标记Lightning-Link?RapidAtto5653xAblabelings(each100-200ugscale)AZK0063C快速Atto565标记Lightning-Link?RapidAtto5651xAblabeling(1scale)AZK0064A快速Atto594标记Lightning-Link?RapidAtto5943xAblabelings(each10-20ugscale)AZK0064B快速Atto594标记Lightning-Link?RapidAtto5943xAblabelings(each100-200ugscale)AZK0064C快速Atto594标记Lightning-Link?RapidAtto5941xAblabeling(1scale)AZK0065A快速罗丹明标记Lightning-Link?RapidRhodamine3xAblabelings(each10-20ugscale)AZK0065B快速罗丹明标记Lightning-Link?RapidRhodamine3xAblabelings(each100-200ugscale)AZK0065C快速罗丹明标记Lightning-Link?RapidRhodamine1xAblabeling(1scale)AZK0066A快速Atto390标记Lightning-Link?RapidAtto3903xAblabelings(each10-20ugscale)AZK0066B快速Atto390标记Lightning-Link?RapidAtto3903xAblabelings(each100-200ugscale)AZK0066C快速Atto390标记Lightning-Link?RapidAtto3901xAblabeling(1scale)AZK0067A快速DyLight350标记Lightning-Link?RapidDyLight3503xAblabelings(each10-20ugscale)AZK0068A快速DyLight350素标记Lightning-Link?RapidDyLight3503xAblabelings(each100-200ugscale)AZK0069A快速DyLight350标记Lightning-Link?RapidDyLight3501xAblabeling(1scale)AZK0070A快速DyLight405标记Lightning-Link?RapidDyLight4053xAblabelings(each10-20ugscale)AZK0070B快速DyLight405标记Lightning-Link?RapidDyLight4053xAblabelings(each100-200ugscale)AZK0070C快速DyLight405标记Lightning-Link?RapidDyLight405
一、探针DNA标记方法
步骤 :
1.10ng~3ugDNA每管,双蒸水定量至总体积15ul
2.沸水水浴10分钟后迅速冰浴
3.加入 5号试剂 2ul , 6号试剂 2ul ,7号试剂 1ul
4.37度1小时~20小时
5.中止反应 加2ul 0.2M EDTA (pH 8.0) 和/或 65度 水浴 10分钟
二、标记效率检测
(一) 试剂配置
Solution Composition Preparation
Washing buffer 0.1 M Maleic acid, 0.15 M NaCl; pH 7.5 (20° C); 0.3% (v/v) Tween 20
Maleic acid buffer 0.1 M Maleic acid, 0.15 M NaCl; adjustwith NaOH (solid) to pH 7.5 (20° C)
Detection buffer 0.1 M Tris-HCl, 0.1 M NaCl, pH 9.5 (20° C)
TE-buffer 10 mM Tris-HCl, 1 mM EDTA, pH 8.0
Blocking stock solution Dissolve Blocking reagent (bottle 10) 10% (w/v) in Maleic
(10 × conc.) acid buffer underconstantly stirring on a heating block(65°C) or heat in a microwave oven,autoclave. The solution remains opaque
Blocking solution Prepare a 1 × working solution by dilutingthe 10 × Blocking solution 1:10in Maleic acid buffer.
Antibody solution Centrifuge Anti-Digoxigenin-AP(vial 8) for 5 min at 10 000 rpm in theoriginal vial prior to each use, and pipet the necessary amount carefully from thesurface. Dilute Anti-
Digoxigenin-AP 1: 5 000 (150 mU/ml) in Blocking solution.
Colorsubstrate Add 40 _l of NBT/BCIP (vial 9) to 2 ml of Detection buffer.
solution Note: Store protected from light!
(二)对照的标记DNA(4号试剂)系列稀释
(三)步骤
1、取以上制备的管2~9各1ul,以及自己标记的探针1ul,点到一小片尼龙膜
2、通过紫外线或者120度半小时使核酸交连到膜上
3、膜置塑料盒中加Maleic acid buffer 20ml ,15~25度轻摇孵育2分钟
4、10ml Blocking solution 孵育30分钟
5、10ml Antibody solution 孵育30分钟
6、10ml Washing buffer 洗2次,每次15分钟
7、10ml Detection buffer中平衡2~5分钟
8、膜在2ml新鲜配制的 Colorsubstrate solution 中避光孵育。显色期间避免摇动
9、中止反应 用TE-buffer或者双蒸水洗5分钟
蛋白酶K预处理
三、样品检测与杂交
(一) 步骤
1、 稀释供试品及阳性对照DNA,点膜
2、 通过紫外线或者120度半小时使核酸交连到膜上
3、 将标记好的探针稀释到约25ng/ml, 煮沸5分钟后迅速冰浴
4、膜放入预热好的预杂交液(3.5ml/100cm2)中,充分混匀,避免起泡
5、倒掉预杂交液,加入杂交液及已变性的探针。杂交温度下至少孵育6小时
四、洗膜
1、2 × SSC, 0.1% SDS , 15-25° C,洗膜2次,每次5分钟。
2、0.5× SSC, 0.1% SDS ,预热至65~68° C,洗膜2次,每次5分钟
五、结果检测
(一) 试剂配制
Solution Composition Preparation
Washing buffer 0.1 M Maleic acid, 0.15 M NaCl; pH 7.5 (20° C); 0.3% (v/v) Tween 20
Maleic acid buffer 0.1 M Maleic acid, 0.15 M NaCl; adjustwith NaOH (solid) to pH 7.5 (20° C)
Detection buffer 0.1 M Tris-HCl, 0.1 M NaCl, pH 9.5 (20° C)
TE-buffer 10 mM Tris-HCl, 1 mM EDTA, pH 8.0
Blocking stock solution Dissolve Blocking reagent (bottle 10) 10% (w/v) in Maleic
(10 × conc.) acid buffer underconstantly stirring on a heating block(65°C) or heat in a microwave oven,autoclave. The solution remains opaque
Blocking solution Prepare a 1 × working solution by dilutingthe 10 × Blocking solution1:10in Maleic acid buffer.
Antibody solution Centrifuge Anti-Digoxigenin-AP(vial 8) for 5 min at 10 000 rpm in theoriginal vial prior to each use, and pipet the necessary amount carefully from thesurface. Dilute Anti-
Digoxigenin-AP 1: 5 000 (150 mU/ml) in Blocking solution.
Colorsubstrate Add 40 _l of NBT/BCIP (vial 9) to 2 ml of Detection buffer.
solution Note: Store protected from light!
(二) 步骤
1、Washing buffer洗膜1~5分钟
2、100ml Blocking solution 孵育30分钟
3、20ml Antibody solution 孵育30分钟
4、100ml Washing buffer 洗2次,每次15分钟
5、20ml Detection buffer中平衡2~5分钟
6、膜在10ml新鲜配制的 Colorsubstrate solution 中避光孵育。显色期间摇动
7、中止反应 用TE-buffer或者双蒸水50ml洗5分钟展开
2、 血浆:EDTA、柠檬酸盐、肝素血浆可用于检测。1000×g离心30分钟去除颗粒。
3、 细胞上清液:1000×g离心10分钟去除颗粒和聚合物。
4、 组织匀浆:将组织加入适量生理盐水捣碎。1000×g离心10分钟,取上清液。
5、 保存:如果样品不立即使用,应将其分成小部分-70℃保存,避免反复冷冻。尽可能的不要使用溶血或高血脂血。如果血清中大量颗粒,检测前先离心或过滤。不要在37℃或更高的温度加热解冻。应在室温下解冻并确保样品均匀地充分解冻。
此IBL试剂盒能用于小鼠血清,EDTA血浆,细胞上清中白介素-6的定量检测 试剂盒成分 1 预包被板: 抗小鼠白介素-6兔子IgG,亲合纯化 96T 2 酶标记抗体: (30倍浓缩)HRP标记抗小鼠白介素-6兔子IgG,亲合纯化 0.4mL x 1 3 标准品: 重组小鼠白介素-6 0.5mL x 2 4 EIA缓冲液: 含1% BSA, 0.05%吐温20 BPS 30mL x 1 5 标记抗体稀释液: 含1% BSA, 0.05%吐温20 BPS 12mL x 1 6 显色剂: TMB底物液 15mL x 1 7 终止液: 1N硫酸 12mL x 1 8 浓缩洗涤液: (40倍浓缩) 含1% BSA, 0.05%吐温20 BPS 50mL x 1 操作说明 1实验所需器材(但试剂盒没有提供) 酶标仪(450nm) 微移液管及其吸嘴 量筒及烧杯 去离子水 冰箱(4°C) 坐标纸(log/log) 吸水纸 试管(用于标准品稀释) 温育箱(37°C ± 1°C) 洗瓶 (用于洗板) 一次性试剂管(用于浓缩酶标记抗体和显色剂)向左转|向右转
探针
ddH2O36.25ul
buffer(瓶3)5ul
核苷酸混合液(含DIG-dUTP)(瓶2)2.5ul
dNTPStockSolution(瓶4)2.5ul
引物F1ul(10pmol/ul)
R1ul(10pmol/ul)
酶(瓶1)0.75ul
DNA1ul(100pg)
程序:35个循环,退火温度55度。
第一次通过探针标记得到了相应的PCR标记产物,亮度和对照相当,片段大小比标记的探针模板大,证明整个过程应该没有问题。
但一个月后重新用相同的体系,相同的程序,只是探针模板的量不同却什么都扩不出来。不知道问题出在哪了!

