| 來源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實(shí)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(cè)或其它適當(dāng)用途時(shí)的一抗稀釋。
建議抗體使用時(shí)的稀釋比例如下(實(shí)際使用時(shí)需根據(jù)抗原水平的高低作適當(dāng)調(diào)整)：
| WB | IP | IF | IHC | ICC | FC | ChIP | ChIP-seq |
| 1:500-1:2000 | - | 1:50-1:200 | - | - | - | - | - |
抗體詳細(xì)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(chǎn)品編號(hào) | 產(chǎn)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(yōu)先推薦4ºC保存，長(zhǎng)期不使用可以考慮-20ºC保存，但凍融可能會(huì)導(dǎo)致出現(xiàn)輕微的渾濁和少量不溶物。
注意事項(xiàng)：
如果本抗體用于Western blot (WB)、免疫熒光(IF)、免疫細(xì)胞化學(xué)(ICC)等實(shí)驗(yàn)，請(qǐng)注意回收使用過的稀釋抗體。回收的抗體通常至少可以重復(fù)使用5-10次。稀釋后的抗體，包括已經(jīng)使用過的稀釋抗體，請(qǐng)4℃保存。
回收后重復(fù)使用的抗體，使用方法同新鮮稀釋的抗體。如果在重復(fù)使用過程中發(fā)現(xiàn)抗體出現(xiàn)輕微混濁現(xiàn)象，可以10,000g離心1-3分鐘，取上清用于后續(xù)檢測(cè)。如果回收的抗體出現(xiàn)明顯的絮狀物或長(zhǎng)霉長(zhǎng)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(xì)胞化學(xué)(ICC)等適當(dāng)用途。如果希望獲得最佳的檢測(cè)效果，請(qǐng)考慮使用上述檢測(cè)專用的一抗稀釋液。
本產(chǎn)品僅限于專業(yè)人員的科學(xué)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(nèi)。
為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。
使用說明：
請(qǐng)根據(jù)抗體的實(shí)際用途選擇相應(yīng)的使用方法。
1. Western檢測(cè)：
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(jīng)過封閉的蛋白膜與稀釋好的一抗4℃緩慢搖動(dòng)過夜或室溫緩慢搖動(dòng)2小時(shí)，確保稀釋的抗體至少能在搖動(dòng)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(xù)使用。
d. 按照Western的實(shí)驗(yàn)步驟進(jìn)行后續(xù)的洗滌、二抗孵育、洗滌和檢測(cè)等操作。
2. 免疫染色：
可以使用生產(chǎn)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(shí)驗(yàn)操作請(qǐng)自行參考適當(dāng)?shù)膒rotocol進(jìn)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. | |
