| 來源 | 用途 | 交叉反應性 | 理論分子量 | 實際分子量 |
| Rabbit | WB, IF, IHC | H, M, R | 24kDa | 29kDa |
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(jù)抗原水平的高低作適當調(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 | - | - | - |
抗體詳細信息如下::
| About this Antibody | |
| Name | HMGB1 Rabbit Polyclonal Antibody (KO Validated) |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | 293T, MCF7, Jurkat, Mouse spleen, Mouse lung, Mouse kidney, Rat lung |
| Cellular location | Cell membrane, Chromosome, Cytoplasm, Endoplasmic reticulum-Golgi intermediate compartment, Endosome, Extracellular side, Nucleus, Peripheral membrane protein, Secreted |
| Customer validation | WB (Human, Mouse); IHC (Mouse, Rat); IF (Mouse) |
| About the Immunogen | |
| Immunogen | A synthetic peptide of human HMGB1 (NP_002119.1). |
| Sequence | SAFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKKAAKLKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEEDEEEE |
| Gene ID | 3146 |
| Swiss Prot | P09429 |
| Synonyms | HMGB1; HMG-1; HMG1; HMG3; SBP-1; high mobility group box 1 |
| Category | Transcription Factors; Epigenetics |
| Background | High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains. All three family members (HMGB1, HMGB2, and HMGB3) contain two HMG box domains and a C-terminal acidic domain. HMGB1 is a widely expressed and highly abundant protein. HMGB2 is widely expressed during embryonic development, but is restricted to lymphoid organs and testis in adult animals. HMGB3 is only expressed during embryogenesis. While expression varies, the biochemical properties of the different family members may be indistinguishable. The HMG box domains facilitate the binding of HMGB proteins to the minor groove of DNA, which results in local bending of the DNA double helix. HMGB proteins are recruited by and help facilitate the assembly of site-specific DNA binding proteins to their cognate binding sites in chromatin. For example, HMGB1 facilitates the binding of Hox proteins, Oct-1, p53, Rel proteins, and steroid hormone receptor proteins to their target gene promoters. In addition to their functions in the nucleus, HMGB proteins play a significant role in extracellular signaling associated with inflammation. HMGB1 is massively released into the extracellular environment during cell necrosis, but not apoptosis. Extracellular HMGB1 "alarms" the innate immune system by acting as a chemoattractant for inflammatory leukocytes, smooth muscle cells, and stem cells, functioning as an immune adjuvant for soluble and particulate antigens, and triggering activation of T cells and dendritic cells. In addition, activated monocytes, macrophages and, dendritic cells also secrete HMGB1, forming a positive feedback loop that results in the release of additional cytokines and neutrophils. Hypoxia has also been shown to cause the release of HMGB1 in the liver, and some studies suggest a role for extracellular HMGB1 in tumor homeostasis. |
包裝清單：
| 產(chǎn)品編號 | 產(chǎn)品名稱 | 包裝 |
| AF5192 | HMGB1 Rabbit Polyclonal Antibody (KO Validated) | 50μl |
| AZ050 | Western一抗稀釋液 | 50ml |
| — | 說明書 | 1份 |
保存條件：
HMGB1 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，但凍融可能會導致出現(xiàn)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。回收的抗體通常至少可以重復使用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(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。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(chǎn)品僅限于專業(yè)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(nèi)。
為了您的安全和健康，請穿實驗服并戴一次性手套操作。
3. 其它實驗操作請自行參考適當?shù)膒rotocol進行。
4. 代表性圖片：
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| Fig. 1. Western blot analysis of extracts of various cell lines, using HMGB1 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 HMGB1 knockout (KO) 293T cells, using HMGB1 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 rat brain using HMGB1 Antibody at dilution of 1:100 (40x lens). | Fig. 4. Immunofluorescence analysis of HeLa cells using HMGB1 antibody at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining. |
