〔Proteintech 解析大全〕如何挑選Internal Control ?
Internal Control 抗體種類很多,其中包含全細胞、胞漿內控、膜內控或核蛋白內控等,如:β- actin、β- tubulin、GAPDH、
Lamin B,以下簡單介紹選擇內控抗體應遵循的原則。
1、樣本種屬來源:首先考慮實驗樣本來源於何物種。
A、哺乳動物的組織或者細胞樣本,通常選擇 β- actin、β- tubulin、GAPDH、Lamin B、Histone H3 等。
B、其他稀少物種來源,可以參照文獻指導或是選擇高度保守houskeeping gene表達的蛋白質相應的抗體作為內控。
2、目標蛋白分子量:
選擇內控抗體時,應該考慮目的蛋白分子量的大小。通常應該保證目的蛋白與內控蛋白分子量相差 5 kDa 以上但勿差距太大。
如:目的蛋白分子量為 45 kDa,此時不適宜選擇 β-actin 作為內控,可以考慮選擇 GAPDH 或者 β- tubulin 作為內控。
3、目標蛋白表達部位:
針對一般蛋白質檢測,GAPDH、β-actin 或 β-tubulin 可以滿足要求,而如果需要檢測細胞器蛋白時,適宜選擇對應細胞器
內控更能體現內部參照的準確性。
如:常用的核內控抗體有 Lamin A、Lamin B、TBP、YY1、HistoneH3。
而對於膜蛋白檢測,常用的內控抗體為 ATP1A1。對於粒線體蛋白的檢測,常用 VDAC1 和 COX IV 作為內控抗體。
≡ 以上原則僅針對通常情況,需要特別注意的是內控的選擇還須考慮實際實驗環境狀況,
如某些細胞或組織由於缺氧、糖尿病等因素會導致 GAPDH 的表達量增高,此種狀況下 GAPDH 不適合做內控;
在涉及細胞增殖相關實驗中,c-Jun 由於自身表達變化就不適合做內控;
在凋亡實驗中,TBP、Lamin 等不適合作為核內控。
在設計實驗時,應考慮上述因素影響並查詢相關文獻,如果在實驗過程中出現內控表達異常狀況,可分析其原因並調整內控選擇。
下方附上小編精心整理的 Internal Control 抗體 推薦列表,大家可以根據上述的內參抗體選擇原則,挑選最適合自己實驗條件的抗體!
Proteintech - Internal Control List
適用範圍 | 名稱 | 貨號 | 分子量 | 抗體種類 | 應用 | 樣本物種 |
全細胞/胞漿 | Vinculin | 66305-1-Ig | 117kDa | Mouse Mono | WB, IHC, IF/ICC, CoIP, ELISA |
human, mouse, rat, pig, zebrafish, hamster |
Alpha Tubulin |
66301-1-Ig |
50-55 kDa |
Mouse Mono | WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA |
human, mouse, rat, rabbit, canine, monkey, chicken, zebrafish, sheep, goat, tick |
|
11224-1-Ig | Rabbit Poly | WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA |
human, mouse, rat, rabbit, monkey, chicken, zebrafish, hamster, goat, p. lividus |
|||
Beta Tubulin |
66240-1-Ig |
50-55 kDa |
Mouse Mono | WB, IHC, IF/ICC, IF-P, IP, ELISA |
human, mouse, rat, pig, zebrafish, nematode, rabbit, canine, monkey, chicken, zebrafish, sheep |
|
10094-1-AP | Rabbit Poly | WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA |
human, mouse, rat, rabbit, chicken, zebrafish, hamster, sheep, goat, fish |
|||
Gamma Tubulin | 66320-1-Ig | 48-55 kDa | Mouse Mono | WB, IHC, IF/ICC, FC (Intra), ELISA |
human, mouse, rat, canine | |
Beta Actin |
42 kDa |
Mouse Mono |
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, ELISA |
human, mouse, rat, pig, rabbit, canine, monkey, chicken, zebrafish, hamster, goat, arabidopsis, golden hamsters, aedes albopictus |
||
Rabbit Poly |
WB, IHC, IF/ICC, IP, CoIP, ELISA, Cell treatment |
human, mouse, rat, canine, monkey, chicken, goat, sheep, yeast, ochotona curzoniae, camelus bactrianus, carp |
||||
GAPDH |
36 kDa |
Mouse Mono |
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA |
human, mouse, rat, pig, zebrafish, yeast, plant, chicken, goat, deer, whiteflies, tick, treeshrew, cynomorium songaricum |
||
Rabbit Poly |
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, RIP, ELISA, Cell treatment |
human, mouse, rat, pig, arabidopsis, cabbage, corn, rice, chicken, goat, sheep, yeast, leech, treeshrew, zebra finches |
||||
細胞核膜 |
Lamin B1 |
66-70 kDa |
Mouse Mono |
WB, IHC, IF/ICC, IF-P, FC (Intra), IP, CoIP, ELISA |
human, mouse, rat, rabbit, chicken, zebrafish, bovine, hamster |
|
Rabbit Poly |
WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ChIP, ELISA |
human, mouse, rat, pig, canine, chicken, bovine, hamster, bombyx mori |
||||
Lamin A/C | 10298-1-AP | 65, 70 kDa | Rabbit Poly | WB, IHC, IF/ICC, FC (Intra), IP, ELISA |
human, mouse, rat, monkey, duck |
|
細胞核 |
YY1 |
65-70 kDa |
Mouse Mono | WB, IHC, IF/ICC, IP, CoIP, ChIP, RIP, ELISA |
human, mouse, rat, monkey, pig |
|
Rabbit Poly | WB, IHC, IF/ICC, IP, CoIP, ChIP, ELISA |
human, mouse, rat, pig, bovine |
||||
TBP |
mouse/rat 33-36 kDa |
Mouse Mono |
WB, IHC, IF/ICC, IP, ELISA | human, mouse, rat, pig | ||
Rabbit Poly |
WB, IHC, IF, IP, ChIP, ELISA | human, mouse, rat, pig, monkey, sheep |
||||
PCNA |
60097-1-Ig |
36-38 kDa |
Mouse Mono | WB, IHC, IF/ICC, IP, CoIP, ELISA |
human, mouse, rat, pig, rabbit, zebrafish |
|
10205-2-AP | Rabbit Poly | WB, IHC, IF/ICC, IF-P, IP, CoIP, ELISA, Cell treatment |
human, mouse, rat, rabbit, chicken, goat, sheep, fish, ducks, medaka embryos |
|||
Histone H3 |
15-17 kDa |
Mouse Mono |
WB, IHC, IF/ICC, FC (Intra), IP, ELISA | human, mouse, rat, pig, rabbit, canine, chicken, zebrafish, hamster, dog, wheat |
||
Rabbit Poly |
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, ELISA | human, mouse, rat, pig, monkey, chicken, goat, fish, arabidopsis, yellow catfish |
||||
粒線體 |
VDAC1/Porin |
66345-1-Ig | 35-37 kDa | Mouse Mono | WB, IHC, IF-P, FC (Intra), IP, CoIP, ELISA | human, mouse, rat, pig |
55259-1-AP | 31 kDa | Rabbit Poly | WB, IHC, IF-P, FC (Intra), CoIP, ELISA, PLA | human, mouse, rat, pig, canine, monkey, chicken, bovine, hamster, duck |
||
VDAC1/2 |
10866-1-AP | 31 kDa | Rabbit Poly | WB, IHC, IF-P, FC (Intra), CoIP, ELISA | human, mouse, rat, pig, rabbit, monkey, zebrafish, pieris rapae |
|
COXIV |
66110-1-Ig |
17-18 kDa |
Mouse Mono | WB, IHC, IF, IP, ELISA | human, mouse, rat | |
11242-1-AP | Rabbit Poly | WB, IHC, IF/ICC, IP, ChIP, ELISA | human, mouse, rat, pig, canine, goat |
|||
COX4I2 | 11463-1-AP | 17-18 kDa | Rabbit Poly | WB, IHC, IF/ICC, IP, ELISA | human, mouse, rat | |
全血/ 血漿/ 血清 |
Transferrin | 66171-1-Ig | 77kDa | Mouse Mono | WB, IHC, IF-P, ELISA | human |
17435-1-AP | Rabbit Poly | WB, IHC, IF/ICC, IP, ELISA | human, mouse, rat, monkey, duck |
|||
Albumin |
66051-1-Ig |
66 kDa |
Mouse Mono | WB, IHC, IF/ICC, IP, ELISA | human, mouse, rat, pig, rabbit |
|
16475-1-AP | Rabbit Poly | WB, IHC, IF/ICC, IF-P, IP, CoIP, ELISA | human, mouse, rat, goat |
|||
膜蛋白 |
ATP1A1 |
100-110 kDa |
Rabbit Poly |
WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA | human, mouse, rat, pig, chicken |
|
ATP1A1-Specific |
100-110 kDa |
Rabbit Poly |
WB, IHC, IF/ICC, FC (Intra), ELISA | human, mouse, rat |
官網連結:Loading Control Antibodies
Proteintech - Internal Control Antibodies 優勢:
身為專注免疫學領域20+年抗體應用專家,Proteintech的內參抗體又有哪些值得你心動的地方呢?
1. 內參種類齊全
Proteintech 提供針對各類樣本的全套內參抗體,
不管是胞漿/全細胞,或是核、膜、粒線體等亞細胞結構,亦或是全血/血漿/血清樣本,均可提供配套的內參抗體;
並且針對同一個目標,還可提供兔多抗、鼠單抗、重組兔單抗等多種選擇,全方位滿足實驗需求。
2. 超高稀釋比
在WB應用中,Proteintech的大部分內參抗體稀釋比可以達到1:10000,部分能達到1:100000,其中GAPDH鼠單抗(貨號:60004-1-Ig)推薦稀釋比更是高達1:50000- 1:500000!
(一支20ul的小包裝,最高可配置10L抗體工作液!(數學好的同學幫忙算算這支抗體可以用多久~>_<))
3. 超多文獻引用
如果說以上的陳述都是小編單方面的陳述,接下來的這個數據絕對客觀、可靠、又令人震撼!
根據第三方平台 CiteAb 資料統計,GAPDH鼠單抗(貨號:60004-1-Ig)目前已被全球科學家引用11909次!也是目前全球被引用最多的一支GAPDH抗體。
除此之外,還有許多 Proteintech - Internal Control 抗體 受到科學家們的肯定,如: GAPDH兔多抗(貨號:10494-1-AP)被引用7103次;
β-Actin鼠單抗(貨號66009-1-Ig)被引用6590次;β-Actin兔多抗(貨號:20536-1-AP)被引用3981次;
α-Tubulin兔多抗(貨號:11224-1-AP)被引用1549次;α-Tubulin鼠單抗(貨號:66031-1-Ig)被引用1109次;
β-Tubulin兔多抗(貨號:10094-1-AP)被引用913次;Lamin B1兔多抗(貨號:12987-1-AP)被引用1183次;
PCNA兔多抗(貨號:10205-2-AP)引用966次;Histone-H3兔多抗(貨號:17168-1-AP )被引用953次......
數萬科學家的選擇,數萬個項目的真實案例驗證,充分佐證了Proteintech抗體的優質可靠!!
看到這裡,對 Proteintech - Internal Control 抗體心動了嗎?
回到標題中的問題:" 如何挑選Internal Control ? "
小編的答案是:" 選 Proteintech 就對啦!"
祝福所有支持 ABreal 倍思特 & Proteintech 的夥伴們,都能實驗計畫事事順利,文獻發表年年如期,科研突破屢創佳績!
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