來源 | 用途 | 交叉反應(yīng)性 | 理論分子量 | 實際分子量 |
Rabbit | WB, IF, IHC | H, M | 20kDa | 23kDa |
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 | CBX3/HP1γ Rabbit Polyclonal Antibody (KO Validated) |
Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
Isotype | IgG |
Purification method | Affinity purification |
Positive samples | HeLa, MCF-7, 293T, A-549, NCI-H460, Mouse brain, Mouse lung, Mouse spleen |
Cellular location | Nucleus |
Customer validation | - |
About the Immunogen | |
Immunogen | Recombinant fusion protein of human CBX3 (NP_009207.2). |
Sequence | MASNKTTLQKMGKKQNGKSKKVEEAEPEEFVVEKVLDRRVVNGKVEYFLKWKGFTDADNTWEPEENLDCPELIEAFLNSQKAGKEKDGTKRKSLSDSESDDSKSKKKRDAADKPRGFARGLDPERIIGATDSSGELMFLMKWKDSDEADLVLAKEANMKCPQIVIAFYEERLTWHSCPEDEAQ |
Gene ID | 11335 |
Swiss Prot | Q13185 |
Synonyms | CBX3; HECH; HP1-GAMMA; HP1Hs-gamma; chromobox 3 |
Category | Histone Methylation; Crosstalk Between Protein Modifications; Epigenetics |
Background | Heterochromatin protein 1 (HP1) is a family of heterochromatic adaptor molecules involved in both gene silencing and higher order chromatin structure. All three HP1 family members (α, β, and γ) are primarily associated with centromeric heterochromatin; however, HP1β and γ also localize to euchromatic sites in the genome. HP1 proteins are approximately 25 kDa in size and contain a conserved amino-terminal chromodomain, followed by a variable hinge region and a conserved carboxy-terminal chromoshadow domain. The chromodomain facilitates binding to histone H3 tri-methylated at Lys9, a histone "mark" closely associated with centromeric heterochromatin. The variable hinge region binds both RNA and DNA in a sequence-independent manner. The chromoshadow domain mediates the dimerization of HP1 proteins, in addition to binding multiple proteins implicated in gene silencing and heterochromatin formation, including the SUV39H histone methyltransferase, the DNMT1 and DNMT3a DNA methyltransferases, and the p150 subunit of chromatin-assembly factor-1 (CAF1). In addition to contributing to heterochromatin formation and propagation, HP1 and SUV39H are also found complexed with retinoblastoma (Rb) and E2F6 proteins, both of which function to repress euchromatic gene transcription in quiescent cells. HP1 proteins are subject to multiple types of post-translational modifications, including phosphorylation, acetylation, methylation, ubiquitination, and sumoylation, suggesting multiple means of regulation. |
包裝清單:
產(chǎn)品編號 | 產(chǎn)品名稱 | 包裝 |
AF5135 | CBX3/HP1γ Rabbit Polyclonal Antibody (KO Validated) | 50μl |
AZ050 | Western一抗稀釋液 | 50ml |
— | 說明書 | 1份 |
保存條件:
CBX3/HP1γ 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(dǎo)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(xué)(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(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。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(xué)(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(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,請穿實驗服并戴一次性手套操作。
使用說明:
請根據(jù)抗體的實際用途選擇相應(yīng)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(jīng)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(xù)使用。
d. 按照Western的實驗步驟進行后續(xù)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(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. 其它實驗操作請自行參考適當?shù)膒rotocol進行。
4. 代表性圖片:
Fig. 1. Western blot analysis of extracts of various cell lines, using CBX3 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 CBX3 knockout (KO) HeLa cells, using CBX3 antibody at 1:500 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 lung cancer using CBX3 antibody at dilution of 1:200 (40x lens). | Fig. 4. Immunofluorescence analysis of U2OS cells using CBX3 antibody at dilution of 1:100. Blue: DAPI for nuclear staining. |