| 流式细胞术及其相关产品的实际应用
流式细胞术是最近发展起来的一项高新生物科学技术。是近代细胞生物学、分子生物学、分子免疫学和单克隆抗体技术、激光技术、计算机技术等学科高度发展,综合应用于临床医学研究及疾病的诊断及治疗的科学结晶。目前已广泛用于肿瘤、血液病、免疫性疾病、艾滋病、器官移植等方面的常规临床检测中,并已成为上述疾病诊断、治疗、疗效判断、预后估计等至关重要甚至唯一的方法。 流式细胞分析术(FCM)能从定量角度对细胞的各种形态学参数、生物学特征、细胞生华成分的组成及含量以及各种功能进行研究。
流式细胞术抗体
Sanquin是一家荷兰公司,是世界上流式细胞术抗体的主要供应商。该公司生产的各种单克隆抗体已被广泛应用于该领域的常规使用。其生产的单克隆抗体具有品种齐全,质量高,效率强的良好信誉。抗体用于分析细胞群体的组成,目前认为,应用此抗体对疾病的诊断、细胞学的研究具有极其重要的意义。
APC通过结合被分解的多肽在免疫反应中起作用,APC会将此多肽片段运至内质网腔,在此处与HLA一型分子结合。抗原多肽片段特定地与由不同的HLA等位基因编码的HLA结合。当它们以足够的亲和力结合,HLA-多肽复合体将被运送到细胞表面。在那儿,此复合物被T细胞的特异性抗原决定簇CD8+识别。这种识别引导T细胞的活化和进一步的免疫反应,同时导致与HLA-多肽复合体融解和脱离。T细胞识别HLA复合体的决定簇就像识别多肽一样,主要是基于以下特性:
T细胞在下列三种情况下会被HLA激活:在接触到同源基因表达的但不相配的HLA会起排斥反应或(一般出现在移植器官中);或接触到异态细胞蛋白作为的抗原多肽(例如肿瘤);或接触到病原体(例如细菌病毒等)。HLA除了与T细胞CD8+分子反应以及细胞内免疫反应,还与其他一种辅助T细胞CD4+分子反应。HLA与CD4+的反应有助于CD8+的活化以及进一步的免疫反应。事实上只有当表达有抗原肽-HLA复合物的APC,与起识别和反应作用的T细胞表面所表达的HLA 分子相同时,二者才能彼此作用而启动免疫应答过程,T细胞表面的TCR在识别抗原肽的过程中,其表面的CD4/CD8分子必须同时识别APC上的HLA-Ⅱ/Ⅰ型分子的Ig样区。T细胞通过产生必要的生长因子和细胞因子在免疫反应中扮演重要角色。CD4+识别HLA二型分子中特殊序列作为抗原 。HLA二型分子位于APC表面,其与被细胞内吞的细胞外蛋白水解反应得到的多肽片段相结合。Heroto,HLA二型分子通过与不同的降糖多肽结合在胞吞途径产生作用。在HLA分子到达细胞表面前,这些降糖多肽随后被水解,然后被由内吞作用得到的多肽所代替。为了优化抗原递呈和T细胞识别,个体内都有多种一型和二型HLA分子,这是由它们各自的亚基所区别的。HLA一型分子由一个恒定的β-微球蛋白亚基和一个变化的α亚基(含有抗原结合位点)。HLA一型分子和二型分子结构上基本一样,但二型分子的αβ亚基大小一样而且两者都含有与抗原结合的特殊位点。体外合成可溶性HLA-多肽复合物被用作多功能复合物在FACS中检测抗原特异性T细胞。
PeliClusterTM Monoclonal Antibodies
可用于流式细胞术的单标记单克隆抗体(FITC-, PE-,PE-Cy5-标记)
包装为1 ml
| Cluster Designation
|
Clone
|
Iso-
type
|
Neat
0.2MG
RUO
|
FITC
100test
CE IVD
|
PE
100test
CE IVD
|
PE-Cy5
100test
CE IVD
|
| CD1a
|
WM35
|
IgG2b
|
M2120
|
M2255
|
M2140
|
|
| CD2
|
CLB-T11/1, 6G4
|
IgG1
|
M1314
|
M1465
|
|
|
| CD3
|
CLB-T3/2, 16A9
|
IgG2a
|
M1306
|
M1466
|
|
|
| CD3
|
UCHT-1
|
IgG1
|
|
|
M1741
|
M2292
|
| CD4
|
CLB-T4/1, 10A12
|
IgG2a
|
M1308
|
|
|
|
| CD4
|
CLB-T4/2, 6D10
|
IgG1
|
|
M1467
|
M1648
|
|
| CD4
|
S3.5
|
IgG2a
|
|
|
|
M1753
|
| CD5
|
CLB-T1/1, 1C12
|
IgG1
|
M1310
|
M1602
|
M1649
|
|
| CD6
|
CLB-B6
|
IgG1
|
M1681
|
|
|
|
| CD7
|
CLB-T-3A1/1, 7F3
|
IgG2a
|
M1339
|
|
|
|
| CD8
|
CLB-T8/4, 4H8
|
IgG2b
|
M1312
|
M1468
|
M1629
|
|
| CD8
|
3B5
|
IgG2a
|
|
|
|
M1744
|
| CD9
|
CLB-thromb/8, 4E1
|
IgG2a
|
M1362
|
M1666
|
|
|
| CD10
|
CLB-CALLA/1, 4F9
|
IgG2a
|
M1337
|
M1603
|
M1692
|
|
| CD11a
|
CLB-LFA-1/2, TB-133
|
IgG2a
|
M1385
|
M1667
|
|
|
| CD11b
|
CLB-mon-gran/1, B2
|
IgM
|
M1386
|
M1668
|
|
|
| CD13
|
CLB-mon-gran/2, Q20
|
IgG2a
|
M1387
|
M1568
|
M1726
|
|
| CD14
|
CLB-mon/1, 8G3
|
IgG2a
|
M1388
|
M1430
|
|
|
| CD14
|
TUK4
|
IgG2a
|
|
|
|
|
| CD15
|
CLB-gran/2, B4
|
IgM
|
M1336
|
M1435
|
|
|
| CD16
|
CLB-FcR gran/1, 5D2
|
IgG2a
|
M1389
|
M1604
|
M1690
|
|
| CD16b
|
CLB-gran/11, 5D7
|
IgG2a
|
M1390
|
|
|
|
| CD18
|
CLB-LFA-1/1, 54
|
IgG1
|
M1344
|
|
|
|
| CD19
|
CLB-B4/1, 11G1
|
IgG1
|
M1345
|
M1436
|
M1588
|
|
| CD19
|
J4.119
|
IgG1
|
|
|
M2285
|
|
| CD19
|
SJ25-C1
|
IgG1
|
|
|
|
|
| CD20
|
NKI-IH4
|
IgG1
|
M1644
|
M1646
|
|
|
| CD21
|
CLB-5D1
|
IgG1
|
M1723
|
M1834
|
|
|
| CD22
|
CLB-B-Ly/1, 6B11
|
IgG1
|
M1391
|
M1437
|
|
|
| CD23
|
Tü1
|
IgG3
|
|
M2108
|
|
|
| CD24
|
CLB-gran-B-ly/1, 1B5
|
IgG1
|
M1361
|
M1605
|
|
|
| CD25
|
CLB-IL2R/1, TB30
|
IgG2b
|
|
M1680
|
|
|
| CD26
|
CLB-22C3
|
IgG1
|
M1696
|
|
|
|
| CD27
|
CLB-CD27/1, 9F4
|
IgG2a
|
M1455
|
M1764
|
|
|
| CD28
|
CLB-CD28/1, 15E8
|
IgG1
|
M1456
|
|
|
|
| CD28
|
KOLT-2
|
IgG1
|
M2241
|
M2180
|
|
|
| CD29
|
2A4
|
IgG1
|
M2129
|
|
|
|
| CD30
|
HSR-4
|
IgG1
|
M2283
|
|
|
|
| CD31
|
CLB-HEC/75
|
IgG1
|
M1536
|
|
|
|
| CD32
|
CIKm5
|
IgG1
|
M2202
|
|
|
|
| CD33
|
CLB-MD33.6
|
IgG1
|
M1585
|
M1606
|
|
|
| CD34
|
ClassI: MD34.2
|
IgG1
|
M1636
|
|
|
|
| CD34
|
ClassII: QBEnd10
|
IgG1
|
|
|
|
|
| CD34
|
ClassIII: 581
|
IgG1
|
|
|
M2284
|
|
| CD35
|
E11
|
IgG1
|
M2122
|
M2136
|
|
|
| CD36
|
CLB-IVC7
|
IgG1
|
M1537
|
M1613
|
|
|
| CD37
|
NMN46
|
IgG2a
|
|
M2139
|
|
|
| CD38
|
CLB-ID5
|
IgM
|
M1642
|
|
|
|
| CD38
|
HIT2
|
IgG1
|
|
|
|
|
| CD40
|
CLB-14G7
|
IgM
|
M1691
|
|
|
|
| CD41
|
CLB-thromb/7, 6C9
|
IgG1
|
M1538
|
M1674
|
|
|
| CD41
|
M148
|
IgG1
|
|
|
M2280
|
|
| CD42b
|
CLB-MB45
|
IgG1
|
M1539
|
M1638
|
M1729
|
|
| CD42b
|
CLB-MB15
|
IgG1
|
M1851
|
|
M1852
|
|
| CD42d
|
CLB-SW16
|
IgG1
|
M1637
|
|
|
|
| CD43
|
DFT-1
|
IgG1
|
M2242
|
|
|
|
| CD44
|
NKI-P2
|
IgG1
|
M1676
|
M1678
|
|
|
| CD45
|
CLB-T200/1, 15D9
|
IgG1
|
M1343
|
M1491
|
|
|
| CD45
|
HI30
|
IgG1
|
|
|
|
|
| CD45R0
|
UCHL1
|
IgG2a
|
M2232
|
M2113
|
|
|
| CD45RA
|
F8-11-13
|
IgG1
|
M2111
|
M2146
|
M2112
|
|
| CD46
|
J4-48
|
IgG1
|
M2148
|
|
|
|
| CD47
|
BRIC126
|
IgG2b
|
M2149
|
|
|
|
| CD48
|
MEM102
|
IgG1
|
M2150
|
|
|
|
| CD49b
|
CLB-thromb/4, 10G11
|
IgG1
|
M1540
|
|
|
|
| CD49c
|
11G5
|
IgG1
|
M2151
|
|
|
|
| CD49d
|
15A8
|
IgG1
|
M2152
|
|
|
|
| CD49e
|
NKI-SAM1
|
IgG2b
|
M1586
|
|
|
|
| CD49f
|
NKI-GoH3
|
IgG2a
|
M1566
|
M1672
|
|
|
| CD51
|
NKI-M9
|
IgG1
|
M1541
|
|
|
|
| CD53
|
MEM53
|
IgG1
|
M2114
|
|
|
|
| CD54
|
15.2
|
IgG1
|
M2115
|
|
|
|
| CD55
|
BRIC110
|
IgG1
|
M2156
|
M2192
|
|
|
| CD56
|
NKI-nbl-1
|
IgG1
|
M1645
|
|
|
|
| CD56
|
B159
|
IgG2a
|
|
|
M2277
|
|
| CD57
|
NC1
|
IgM
|
M2204
|
|
|
|
| CD58
|
BRIC5
|
IgG2a
|
M2157
|
|
|
|
| CD59
|
MEM43
|
IgG2a
|
M2121
|
|
|
|
| CD61
|
CLB-thromb/1, C17
|
IgG1
|
M1355
|
M1592
|
|
|
| CD62P
|
CLB-thromb/6, C2
|
IgG1
|
M1543
|
|
|
|
| CD62P
|
AK6
|
IgG1
|
|
|
|
|
| CD63
|
CLB-gran/12, 435
|
IgG1
|
M1544
|
|
M1698
|
|
| CD64
|
10.1
|
IgG1
|
M2159
|
|
|
|
| CD65
|
88H7
|
IgM
|
M1719
|
|
|
|
| CD66b
|
CLB-B13.9
|
IgG1
|
M1546
|
M1594
|
|
|
| CD66acde
|
CLB-gran/10, IH4Fc
|
IgG1
|
M1545
|
M1643
|
M1693
|
|
| CD69
|
TP1.55.3
|
IgG2b
|
|
|
|
|
| CD70
|
CLB-2F2
|
IgG1
|
M1705
|
|
|
|
| CD71
|
RVS10
|
IgG1
|
M2160
|
M2162
|
M2119
|
|
| CD74
|
BU45
|
IgG1
|
M2163
|
|
|
|
| CD80
|
CLB-DAL1
|
IgG1
|
M1708
|
|
|
|
| CD95
|
FAS18
|
IgG2a
|
|
|
|
|
| CD95
|
FAS19
|
IgG1
|
M1715
|
|
|
|
| CD103
|
2G5
|
IgG2a
|
M1721
|
M1722
|
| CD105
|
CLB-HEC-19
|
IgG1
|
M1765
|
|
|
|
| CD122
|
MIK-b1
|
IgG2a
|
M2172
|
|
|
|
| CD138
|
CLB-1D4
|
IgG1
|
M1663
|
|
M2287
|
|
| CD235a
|
CLB-AME-1
|
IgG1
|
M1365
|
|
|
|
单标记对照抗体:
| Controls
|
Iso-type
|
Neat
|
FITC-label
|
PE-label
|
PE-Cy5
|
|
IgG1 |
IgG1 |
M1451 |
M1453 |
M1628 |
|
|
IgG2a |
IgG2 |
M1452 |
M1454 |
M1697 |
M1748 |
| IgG2b
|
IgG2b
|
M2173
|
M2190
|
M2230
|
|
非CD抗体/颗粒酶抗体
| Antibody
|
Clone
|
Iso-type
|
Neat
|
FITC
-label
|
PE
-label
|
Biotin
-label
|
| Ery-H/1
|
CLB-7C11
|
IgM
|
M1392
|
|
|
|
| MPO
|
CLB-MPO-1/1, 7.17
|
IgG1
|
M1464
|
M1703
|
|
|
| LTF
|
CLB-13.17
|
IgG1
|
M1700
|
|
M1731
|
|
| HLA-DR
|
CLB-HLA-DR, 1E5
|
IgG1
|
M1338
|
M1607
|
|
|
| ANCA
|
CLB-12.8
|
IgG1
|
M1574
|
|
|
|
| GA6
|
Granzyme A
|
IgG1
|
M1791
|
|
|
|
| GA28
|
Granzyme A
|
IgG1
|
M1766
|
|
|
M1798
|
| GA29
|
Granzyme A
|
IgG1
|
M1767
|
|
|
|
| GB7
|
Granzyme B
|
IgG2a
|
M1754
|
|
|
|
| GB10
|
Granzyme B
|
IgG1
|
M1755
|
|
|
M1793
|
| GB11
|
Granzyme B
|
IgG1
|
M1756
|
|
M2289
|
M1794
|
| GB12
|
Granzyme B
|
IgG1
|
M1757
|
|
|
M1759
|
PeliDuo™ Monoclonal Antibodies Set
用于流式细胞的双标单克隆抗体(FITC / PE标记)
包装为1 ml,可以进行50检测
| Antibody combinations 50 tests
|
Clone
|
Iso-type
|
Order number
CE IVD
|
|
CD2 FITC / CD19 PE |
CLB-T11/1 / CLB-B4/1 |
IgG1/IgG1
|
M1624
|
|
CD3 FITC / CD4 PE |
CLB-T3/2 / CLB-T4/2 |
IgG2a/IgG1
|
M2247
|
|
CD3 FITC / CD8 PE |
CLB-T3/2 / CLB-T8/4 |
IgG2a/IgG2b1
|
M1598
|
|
CD3 FITC / CD16 PE |
CLB-T3/2 / CLB-FcRgran/1 |
IgG2a/IgG2b
|
M1724
|
|
CD3 FITC / CD19 PE |
CLB-T3/2 / CLB-B4/1 |
IgG2a/IgG1
|
M1699
|
|
CD4 FITC / CD8 PE |
CLB-T4/2 / CLB-T8/4 |
IgG1/IgG2b
|
M1599
|
|
CD14 FITC / CD45 PE |
CLB-mon/1 / CLB-T200/1 |
IgG2a/IgG1
|
M1626
|
|
CD45 FITC / CD14 PE |
CLB-T200/1 / CLB-mon/1 |
IgG1/IgG1
|
M2248
|
双标记对照抗体:
|
Controls |
Iso-type |
Order number |
|
IgG1-FITC/IgG1-PE |
IgG1/ IgG1 |
M1627 |
|
IgG2a-FITC/IgG1-PE |
IgG2a/ IgG1 |
M2251 |
PeliTrio™ Monoclonal Antibodies
用于流式细胞的三标单克隆抗体(FITC / PE / PE-Cy5标记)
包装:1ml 可做100检测 (including lysing solution*)
| Antibody combinations 100 tests
|
Clone
|
Order number CE IVD
|
| CD4 FITC / CD8 PE / CD3 PE-Cy5
|
CLB-T4/2 / CLB-T8/4 / UCHT-1S4.1
|
M1825.s
|
| CD16 FITC / CD19PE / CD3 PE-Cy5
|
3G8 / 4G7 / UCHT-1
|
M1799.s
|
|
三标记对照抗体 |
|
control IgG2a FITC / IgG2a PE / IgG2a PE-Cy5 |
M1826.s
|
PeliClusterTM Cell-Stimulating Antibodies
用于T细胞的增殖和功能研究的淋巴细胞刺激的单克隆抗体
包装:0.1 ml (2 mg/mL)
| Antibody
|
Order number
|
Isotype
|
Clone name
|
Application
|
| CD2
|
M1651
|
IgG1
|
CLB-T11.1/, 6G4
|
T cell stimulation
|
| CD2
|
M1652
|
IgG1
|
CLB-T11.2/, 4B2
|
T cell stimulation
|
| CD2
|
M1653
|
IgG1
|
CLB-HIK27
|
T cell stimulation
|
| CD3
|
M1654
|
IgE
|
CLB-T3/4E, 1XE
|
T cell stimulation
|
| CD3
|
M1665
|
IgG1
|
CLB-T3/2, 16A9
|
T cell stimulation
|
| CD28
|
M1650
|
IgG1
|
CLB-CD28/1, 15E8
|
T cell co-stimulation
|
PeliTrio monoclonal antibodies for use in ORTHO ImmunoCount system
PeliTrio单克隆抗体用于ORTHO免疫计数
100 tests(Including lysing solution)
|
Antibody combinations |
Order number |
|
CD4 FITC / CD8 PE / CD3 PE-Cy5 |
M1825.s |
|
CD16 FITC / CD19 PE / CD3 PE-Cy5 |
M1799.s |
|
Isotype control |
|
|
IgG2a FITC / IgG2a PE / IgG2a PE-Cy5 |
|
|
Lysing solution |
|
|
Lysing solution (300 tests) |
M7101.6 |
MHC四聚体(人、小鼠)- PeliMersTM
Sanquin是世界上MHC四聚体的主要供应商。除提供重组MHC外,Sanquin还为客户合成 MHC-Peptide化合物的专业公司,它能根据客户所提供的肽序列,为客户合成特殊的MHC-Peptide化合物。
MHC四聚体是第一种可在流式中鉴别抗原特异性T细胞的试剂,其革命性改进了免疫学研究。该技术逐渐被认为是定量T细胞免疫反应的金标准。其最初的文献(Phenotypic Analysis of Antigen-Specific T Lymphocytes; Altman, John et. al. Science 1996 274: 94-96.)已被引用超过500次。用MHC-肽四聚体可在流式细胞仪上从大量细胞中检测和分离抗原特异性T细胞。可用于分析新鲜或冰冻细胞。不仅节约时间,而且更精确。细胞表型分析可以与细胞内细胞因子检测同时分析,使您在一次分析中了解细胞的特性和激活状态。
Sanquin提供各种包装PeliMers MHC四聚体
(10 Test、50 Test、250 Test)
人
|
Human |
Epitope |
Allele |
Description |
Order code |
|
Virus antigens |
|
|
|
|
|
|
FLU |
|
|
CTELKLSDY |
A*0101 |
PeliMer A1/FLU PE |
M2425CTELK |
|
VSDGGPNLY |
A*0101 |
PeliMer A1/FLU PE |
M2425VSDGG |
|
|
|
HIV |
|
|
SLYNTVATL |
A*0201 |
PeliMer A2/HIV-gag PE |
M2427SLYNT |
|
SLYNTVATL |
A*0201 |
PeliMer A2/HIV-gag APC |
M2428SLYNT |
|
RLRPGGKKK |
A*0301 |
PeliMer A3/HIV-gag PE |
M2429RLRPG |
|
RLRPGGKKK |
A*0301 |
PeliMer A3/HIV-gag APC |
M2430RLRPG |
|
IVLPEKDSW |
B*5701 |
PeliMer B57/HIV-pol PE |
M2439IVLPE |
|
KAFSPEVIPMF |
B*5701 |
PeliMer B57/HIV-gag PE |
M2439KAFSP |
|
|
|
CMV |
|
|
NLVPMVATV |
A*0201 |
PeliMer A2/CMV (pp65) |
M2427NLVPM |
|
|
|
EBV |
|
|
RLRAEAQVK |
A*0301 |
PeliMer A3/EBV PE |
M2429RLRAE |
|
KIRLRPGGK |
A*0301 |
PeliMer A3/EBV APC |
M2430KIRLR |
|
IVTDFSVIK |
A*1101 |
PeliMer A11/EBV PE |
M2431IVTDF |
|
AVFDRKSDAK |
A*1101 |
PeliMer A11/EBV PE |
M2431AVFDR |
|
AVFDRKSDAK |
A*1101 |
PeliMer A11/EBVAPC |
M2432AVFDR |
|
RPPIFIRRL |
B*0702 |
PeliMer B7/EBV PE |
M2433RPPIF |
|
RPPIFIRRL |
B*0702 |
PeliMer B7/EBV APC |
M2434RPPIF |
|
RAKFKQLL |
B*0801 |
PeliMer B8/EBV PE |
M2435RAKFK |
|
EPLPQGQLTAY |
B*3501 |
PeliMer B35/BZLF1 PE |
M2437EPLPQ |
|
HPVGEADYFEY |
B*3501 |
PeliMer B35/EBNA1 PE |
M2437HPVGE |
|
|
|
RSV |
|
|
* |
A*0201 |
PeliMer A2/RSV APC |
M2428O2927 |
|
* |
B*0702 |
PeliMer B7/RSV APC |
M2434O2936 |
|
TPKGPSLRV |
B*0702 |
PeliMer B7/RSV APC |
M2434TPKGP |
|
|
|
HBV |
|
|
FLLSLGIHL |
A*0201 |
PeliMer A2/HBV-pol PE |
M2427FLLSL |
|
FLPSDFFPSV |
A*0201 |
PeliMer A2/ HBV-core PE |
M2427FLPSD |
|
WLSLLWPFV |
A*0201 |
PeliMer A2/ HBV-env PE |
M2427WLSLL |
|
|
|
HPV |
|
|
KLPQLCTEL |
A*0201 |
PeliMer A2/HPV PE |
M2427KLPQL |
|
Tumour antigens |
|
|
|
|
|
|
TUMOUR |
|
|
YMDGTMSQV |
A*0201 |
PeliMer A2/ Tyrosinase PE |
M2427YMDGT |
|
RMFPNAPYL |
A*0201 |
PeliMer A2/ WT-1 PE |
M2427RMFPN |
小鼠
|
Mu
r
I
N
e |
Epitope |
Allele |
Description |
Order code |
|
SIINFEKL |
H2-Kb |
PeliMer H2-Kb / OVA APC |
M2711SIINF |
|
SVYDFFVWL |
H2-Kb |
PeliMer H2-Kb /TRP-2 APC |
M2711SVYDF |
|
ASNENMDAM |
H2-Db |
PeliMer H2-Db /F5 PE |
M2712ASNEN |
|
KCSRNRQYL |
H2-Db |
PeliMer H2-Db / HY PE |
M2712KCSRN |
|
RAHYNIVTF |
H2-Db |
PeliMer H2-Db / PE |
M2712RAHYN |
|
ASNENMDAM |
H2-Db |
PeliMer H2-Db / F5 APC |
M2713ASNEN |
|
KCSRNRQYL |
H2-Db |
PeliMer H2-Db / HY APC |
M2713KCSRN | |