來源 | 用途 | 交叉反應性 | 理論分子量 | 實際分子量 |
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 | 1:50-1:100 | 1:50-1:200 | 1:50-1:200 | 1:50-1:200 | - | - | - |
抗體詳細信息如下::
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:
產品編號 | 產品名稱 | 包裝 |
AF5123 | Beclin 1 Rabbit Polyclonal Antibody (KO Validated) | 50μl |
AZ050 | Western一抗稀釋液 | 50ml |
— | 說明書 | 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。
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 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. |