來源
用途
交叉反應性
理論分子量
實際分子量
Rabbit
WB, IF
H, M, R
71/79kDa
80kDa
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk, Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀釋液，可以用于Western檢測或其它適當用途時的一抗稀釋。
建議抗體使用時的稀釋比例如下(實際使用時需根據抗原水平的高低作適當調整)：
WB
IP
IF
IHC
ICC
FC
ChIP
ChIP-seq
1:500-1:2000
-
1:50-1:200
-
-
-
-
-
抗體詳細信息如下::
About this Antibody
Name
CAPN2 Rabbit Polyclonal Antibody (KO Validated)
Category
Rabbit Polyclonal Antibody (pAb); Primary antibody
Isotype
IgG
Purification method
Affinity purification
Positive samples
H460, A-549, HeLa, Mouse kidney, Mouse lung
Cellular location
Cell membrane, Cytoplasm
Customer validation
WB (Pig)
About the Immunogen
Immunogen
Recombinant fusion protein of human CAPN2 (NP_001739.2).
Sequence
SDTYKKWKLTKMDGNWRRGSTAGGCRNYPNTFWMNPQYLIKLEEEDEDEEDGESGCTFLVGLIQKHRRRQRKMGEDMHTIGFGIYEVPEELSGQTNIHLSKNFFLTNRARERSDTFINLREVLNRFKLPPGEYILVPSTFEPNKDGDFCIRVFSEKKADYQAVDDEIEANLEEFDISEDDIDDGFRRLFAQLAGEDAEISAFELQTILRRVLAKRQDIKSDGFSIETCKIMVDMLDSDGSGKLGLKEFYILWTKIQKYQKIYREIDVDRSGTMNSYEMRKALEEAGFKMPCQLHQVIVARFADDQLIIDFDNFVRCLVRLETLFKIFKQLD
Gene ID
824
Swiss Prot
P17655
Synonyms
CAPN2; CANP2; CANPL2; CANPml; mCANP; calpain 2
Category
Adherens Junction Dynamics; PI3K/Akt Signaling; mTOR Signaling; Breast Cancer
Background
Calpain is a calcium-dependent thiol proteinase that is functionally active as a heterodimer composed of a small regulatory subunit and one of at least two large catalytic subunits (calpain 1 or calpain 2). In vitro, calpain 1 (mu-calpain) requires micromolar levels of calcium, while calpain 2 (M-calpain) requires millimolar levels of calcium for activation. The regulation of calpain in vivo is the subject of many current studies, which suggest that proteolytic activity is regulated post-transcriptionally by mechanisms such as calcium requirements, subcellular localization of the heterodimer, phosphorylation via the EGFR-Erk signaling cascade, endogenous inhibitors (calpastatin) and autoproteolytic cleavage. Calpastatin negatively regulates autoproteolytic cleavage of calpain 1 between Gly27 and Leu28. Calpain influences cell migration by modifying rather than degrading its substrates responsible for cell adhesion and cytoskeletal arrangement. Control of calpain activity has caught the attention of drug development since limiting its activity could mute invasiveness of tumors or chronic inflammation.
包裝清單：
產品編號
產品名稱
包裝
AF5129
CAPN2 Rabbit Polyclonal Antibody (KO Validated)
50μl
AZ050
Western一抗稀釋液
50ml
—
說明書
1份
保存條件：
CAPN2 Rabbit Polyclonal Antibody (KO Validated) -20oC保存，Western一抗稀釋液-20oC或4oC保存，一年有效。Western一抗稀釋液優先推薦4oC保存，長期不使用可以考慮-20oC保存，但凍融可能會導致出現輕微的渾濁和少量不溶物。
注意事項：
如果本抗體用于Western blot (WB)、免疫熒光(IF)、免疫細胞化學(ICC)等實驗，請注意回收使用過的稀釋抗體?；厥盏目贵w通常至少可以重復使用5-10次。稀釋后的抗體，包括已經使用過的稀釋抗體，請4℃保存。
回收后重復使用的抗體，使用方法同新鮮稀釋的抗體。如果在重復使用過程中發現抗體出現輕微混濁現象，可以10,000g離心1-3分鐘，取上清用于后續檢測。如果回收的抗體出現明顯的絮狀物或長霉長菌等情況，則可以考慮廢棄該抗體。
提供的Western一抗稀釋液也可以用于免疫熒光(IF)、免疫組化(IHC)、免疫細胞化學(ICC)等適當用途。如果希望獲得最佳的檢測效果，請考慮使用上述檢測專用的一抗稀釋液。
本產品僅限于專業人員的科學研究用，不得用于臨床診斷或治療，不得用于食品或藥品，不得存放于普通住宅內。
為了您的安全和健康，請穿實驗服并戴一次性手套操作。
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使用說明：
請根據抗體的實際用途選擇相應的使用方法。
1. Western檢測：
a.?按照推薦的稀釋比例用提供的Western一抗稀釋液稀釋抗體。
b.?把經過封閉的蛋白膜與稀釋好的一抗4℃緩慢搖動過夜或室溫緩慢搖動2小時，確保稀釋的抗體至少能在搖動的瞬間覆蓋蛋白膜。
c.?回收稀釋的一抗，4℃保存以備下次繼續使用。
d.?按照Western的實驗步驟進行后續的洗滌、二抗孵育、洗滌和檢測等操作。
2. 免疫染色：
可以使用生產的免疫染色一抗稀釋液(P0103)稀釋抗體，使用后注意回收稀釋好的一抗，
3. 其它實驗操作請自行參考適當的protocol進行。
4. 代表性圖片：
Fig. 1. Western blot analysis of extracts of various cell lines, using CAPN2 antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25μg per lane. Blocking buffer: QuickBlock? Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s.
Fig. 2. Western blot analysis of extracts from normal (control) and CAPN2 knockout (KO) 293T cells, using CAPN2 antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25μg per lane. Blocking buffer: QuickBlock? Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s.
Fig. 3. Immunofluorescence analysis of U2OS cells using CAPN2 antibody at dilution of 1:100. Blue: DAPI for nuclear staining.
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