| 来源 | 用途 | 交叉反应性 | 理论分子量 | 实际分子量 |
| Rabbit | WB, IP, IF, IHC | H, M, R | 38/47kDa | 62kDa |
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一抗稀释液，可以用于Western检测或其它适当用途时的一抗稀释。
建议抗体使用时的稀释比例如下(实际使用时需根据抗原水平的高低作适当调整)：
| WB | IP | IF | IHC | ICC | FC | ChIP | ChIP-seq |
| 1:500-1:2000 | 1:20-1:50 | 1:50-1:100 | 1:50-1:100 | 1:50-1:100 | - | - | - |
抗体详细信息如下::
| About this Antibody | |
| Name | SQSTM1/p62 Rabbit Polyclonal Antibody (KO Validated) |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | 293T, HeLa, U-87MG |
| Cellular location | Cytoplasm, Cytoplasmic vesicle, Endoplasmic reticulum, Late endosome, Lysosome, Nucleus, P-body, autophagosome |
| Customer validation | WB (Rat) |
| About the Immunogen | |
| Immunogen | Recombinant fusion protein of human SQSTM1 / p62 (NP_001135770.1). |
| Sequence | MAMSYVKDDIFRIYIKEKKECRRDHRPPCAQEAPRNMVHPNVICDGCNGPVVGTRYKCSVCPDYDLCSVCEGKGLHRGHTKLAFPSPFGHLSEGFSHSRWLRKVKHGHFGWPGWEMGPPGNWSPRPPRAGEARPGPTAESASGPSEDPSVNFLKNVGESVAAALSPLGIEVDIDVEHGGKRSRLTPVSPESSSTEEKSSSQPSSCCSDPSKPGGNVEGATQSLAEQMRKIALESEGRPEEQMESDNCSGGDDDWTHLSSKEVDPSTGELQSLQMPESEGPSSLDPSQEGPTGLKEAALYPHLPPEADPRLIESLSQMLSMGFSDEGGWLTRLLQTKNYDIGAALDTIQYSKHPPPL |
| Gene ID | 8878 |
| Swiss Prot | Q13501 |
| Synonyms | SQSTM1; A170; DMRV; FTDALS3; NADGP; OSIL; PDB3; ZIP3; p60; p62; p62B; sequestosome-1 |
| Category | Ubiquitin/Proteasome; Autophagy Signaling; NF-κB Signaling |
| Background | Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy. It was first identified as a protein that binds to the SH2 domain of p56Lck and independently found to interact with PKCζ. SQSTM1 was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome. Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway. Protein aggregates formed by SQSTM1 can be degraded by the autophagosome. SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome. Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress. Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity. |
包装清单：
| 产品编号 | 产品名称 | 包装 |
| AF5312 | SQSTM1/p62 Rabbit Polyclonal Antibody (KO Validated) | 50μl |
| AZ050 | Western一抗稀释液 | 50ml |
| — | 说明书 | 1份 |
保存条件：
SQSTM1/p62 Rabbit Polyclonal Antibody (KO Validated) -20ºC保存，Western一抗稀释液-20ºC或4ºC保存，一年有效。Western一抗稀释液优先推荐4ºC保存，长期不使用可以考虑-20ºC保存，但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项：
如果本抗体用于Western blot (WB)、免疫荧光(IF)、免疫细胞化学(ICC)等实验，请注意回收使用过的稀释抗体。回收的抗体通常至少可以重复使用5-10次。稀释后的抗体，包括已经使用过的稀释抗体，请4℃保存。
回收后重复使用的抗体，使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象，可以10,000g离心1-3分钟，取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况，则可以考虑废弃该抗体。
提供的Western一抗稀释液也可以用于免疫荧光(IF)、免疫组化(IHC)、免疫细胞化学(ICC)等适当用途。如果希望获得最佳的检测效果，请考虑使用上述检测专用的一抗稀释液。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明：
请根据抗体的实际用途选择相应的使用方法。
1. Western检测：
a. 按照推荐的稀释比例用碧云天提供的Western一抗稀释液稀释抗体。
b. 把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜或室温缓慢摇动2小时，确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
c. 回收稀释的一抗，4℃保存以备下次继续使用。
d. 按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。具体操作可以参考如下网页：
http://www.beyotime.com/western.htm
2. 免疫染色：
可以使用碧云天生产的免疫染色一抗稀释液(P0103)稀释抗体，使用后注意回收稀释好的一抗，具体操作可以参考如下网页：http://www.beyotime.com/support/immunol-staining.htm
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片：
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| Fig. 1. Western blot analysis of extracts of various cell lines, using SQSTM1 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 SQSTM1 / p62 knockout (KO) 293T cells, using SQSTM1 / p62 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. |
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| Fig. 3. Immunohistochemistry of paraffin-embedded human esophagus using SQSTM1 / p62 antibody at dilution of 1:100 (40x lens). | Fig. 4. Immunofluorescence analysis of U2OS cells using SQSTM1 / p62 antibody at dilution of 1:100. Blue: DAPI for nuclear staining. |
