| 來(lái)源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實(shí)際分子量 |
| Rabbit | WB, IP, IF, IHC | H, M, R | 51kDa | 60kDa |
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 | 1:50-1:100 | 1:50-1:200 | 1:50-1:200 | 1:50-1:200 | - | - | - |
抗體詳細(xì)信息如下::
| About this Antibody | |
| Name | Beclin 1 Rabbit Polyclonal Antibody (KO Validated) |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | U-87MG, HeLa, MCF7, Mouse spleen, Rat ovary, Rat lung |
| Cellular location | Cytoplasm, Cytoplasmic vesicle, Endoplasmic reticulum membrane, Endosome, Endosome membrane, Golgi apparatus, Mitochondrion, Mitochondrion membrane, Nucleus, Peripheral membrane protein, autophagosome, trans-Golgi network membrane |
| Customer validation | WB (Mouse, Pig, Rat, Human); IHC (Mouse); IHC (Mouse); WB (Mouse) |
| About the Immunogen | |
| Immunogen | Recombinant fusion protein of human Beclin 1 (NP_003757.1). |
| Sequence | MEGSKTSNNSTMQVSFVCQRCSQPLKLDTSFKILDRVTIQELTAPLLTTAQAKPGETQEEETNSGEEPFIETPRQDGVSRRFIPPARMMSTESANSFTLIGEASDGGTMENLSRRLKVTGDLFDIMSGQTDVDHPLCEECTDTLLDQLDTQLNVTENECQNYKRCLEILEQMNEDDSEQLQMELKELALEEERLIQELEDVEKNRKIVAENLEKVQAEAERLDQEEAQYQREYSEFKRQQLELDDELKSVENQMRYAQTQLDKLKKTNVFNATFHIWHSG |
| Gene ID | 8678 |
| Swiss Prot | Q14457 |
| Synonyms | BECN1; ATG6; VPS30; beclin1; beclin-1 |
| Category | Autophagy Signaling |
| Background | Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of proteins activated in response to nutrient deprivation and in neurodegenerative conditions. One of the proteins critical to this process is Beclin-1, the mammalian orthologue of the yeast autophagy protein Apg6/Vps30. Beclin-1 can complement defects in yeast autophagy caused by loss of Apg6 and can also stimulate autophagy when overexpressed in mammalian cells. Mammalian Beclin-1 was originally isolated in a yeast two-hybrid screen for Bcl-2 interacting proteins and has been shown to interact with Bcl-2 and Bcl-xL, but not with Bax or Bak. While Beclin-1 is generally ubiquitously expressed, research studies have shown it is monoallelically deleted in 40-75% of sporadic human breast and ovarian cancers. Beclin-1 is localized within cytoplasmic structures including the mitochondria, although overexpression of Beclin-1 reveals some nuclear staining and CRM1-dependent nuclear export. Investigators have demonstrated that Beclin-1-/- mice die early in embryogenesis and Beclin-1-/+ mice have a high incidence of spontaneous tumors. Stem cells from the null mice demonstrate an altered autophagic response, although responses to apoptosis appeared normal. Researchers have also found that overexpression of Beclin-1 in virally infected neurons in vivo resulted in significant protection against Sindbis virus-induced disease and neuronal apoptosis. |
包裝清單：
| 產(chǎn)品編號(hào) | 產(chǎn)品名稱(chēng) | 包裝 |
| AF5123 | Beclin 1 Rabbit Polyclonal Antibody (KO Validated) | 50μl |
| AZ050 | Western一抗稀釋液 | 50ml |
| — | 說(shuō)明書(shū) | 1份 |
保存條件：
Beclin 1 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(guò)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(guò)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(guò)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(zhuān)用的一抗稀釋液。
本產(chǎn)品僅限于專(zhuān)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(shuō)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(guò)封閉的蛋白膜與稀釋好的一抗4℃緩慢搖動(dòng)過(guò)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 Beclin 1 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 Beclin 1 knockout (KO) 293T cells, using Beclin 1 antibody at 1:3000 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. Immunohistochemistry of paraffin-embedded human breast cancer using Beclin 1 antibody at dilution of 1:100 (40x lens). | Fig. 4. Immunofluorescence analysis of HeLa cells using Beclin 1 antibody at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining. |
