| 來源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實(shí)際分子量 |
| Rabbit | WB, IF, IHC | H, M, R, Pg | 14kDa | 14/16kDa |
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 | 1:50-1:200 | 1:50-1:200 | - | - | - |
抗體詳細(xì)信息如下::
| About this Antibody | |
| Name | LC3B Rabbit Polyclonal Antibody (KO Validated) |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | C6, HeLa, NIH/3T3, Pig liver |
| Cellular location | Cytoplasm, Cytoplasmic vesicle, Endomembrane system, Lipid-anchor, autophagosome, autophagosome membrane, cytoskeleton |
| Customer validation | WB (Mouse); IHC (Mouse) |
| About the Immunogen | |
| Immunogen | A synthetic peptide of human LC3B |
| Sequence | - |
| Gene ID | 81631 |
| Swiss Prot | Q9GZQ8 |
| Synonyms | MAP1LC3B; ATG8F; LC3B; MAP1A/1BLC3; MAP1LC3B-a; microtubule associated protein 1 light chain 3 beta |
| Category | Organelle Markers; Autophagy Signaling |
| Background | Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection, and cancer. Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy. Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy. Cleavage of LC3 at the carboxy terminus immediately following synthesis yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II through lipidation by a ubiquitin-like system involving Atg7 and Atg3 that allows for LC3 to become associated with autophagic vesicles. The presence of LC3 in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy. |
包裝清單：
| 產(chǎn)品編號(hào) | 產(chǎn)品名稱 | 包裝 |
| AF5225 | LC3B Rabbit Polyclonal Antibody (KO Validated) | 50μl |
| AZ050 | Western一抗稀釋液 | 50ml |
| — | 說明書 | 1份 |
保存條件：
LC3B 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�；厥盏目贵w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(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 LC3B antibody at 1:1000 dilution. C6 cells were treated by Chloroquine for 20 hours. Hela cells were treated by Chloroquine for 20 hours. NIH/3T3 cells were treated by Chloroquine for 20 hours. 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 of pig liver, using LC3B antibody at 1:10000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). |
 |  |
| Fig. 3. Immunohistochemistry of paraffin-embedded rat heart using LC3B antibody at dilution of 1:100 (40x lens). | Fig. 4. Immunofluorescence analysis of NIH/3T3 cells using LC3B antibody at dilution of 1:100. NIH/3T3 cells were treated by Chloroquine (50 μM) for 20 hours(left). Blue: DAPI for nuclear staining. |
