來源 | 用途 | 交叉反應性 | 理論分子量 | 實際分子量 |
Rabbit | WB, IF | H, M, R | 250kDa | 320kDa |
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 | - | 1:50-1:200 | - | - | - | - | - |
抗體詳細信息如下:
About this Antibody | |
Name | CSPG4/NG2/MCSP Rabbit Polyclonal Antibody (KO Validated) |
Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
Isotype | IgG |
Purification method | Affinity purification |
Positive samples | A-375, HeLa |
Cellular location | Apical cell membrane, Cell projection, Extracellular side, Single-pass type I membrane protein, lamellipodium membrane |
Customer validation | - |
About the Immunogen | |
Immunogen | Recombinant fusion protein of human CSPG4 (NP_001888.2). |
Sequence | LEVPQALGRSSLSQQQLRVVSDREEPEAAYRLIQGPQYGHLLVGGRPTSAFSQFQIDQGEVVFAFTNFSSSHDHFRVLALARGVNASAVVNVTVRALLHVWAGGPWPQGATLRLDPTVLDAGELANRTGSVPRFRLLEGPRHGRVVRVPRARTEPGGSQLVEQFTQQDLEDGRLGLEVGRPEGRAPGPAGDSLTLELWAQGVPPAVASLDFATEPYNAARPYSVALLSVPEAARTEAGKPESSTPTGEPGPMASSPEPAVA |
Gene ID | 1464 |
Swiss Prot | Q6UVK1 |
Synonyms | CSPG4; HMW-MAA; MCSP; MCSPG; MEL-CSPG; MSK16; NG2; chondroitin sulfate proteoglycan 4 |
Category | Neuronal Signaling |
Background | The chondroitin sulfate proteoglycan CSPG4 (chondroitin sulfate proteoglycan 4), also named as NG2, is a type I membrane protein expressed by subpopulations of glia including oligodendroglial precursor cells and a variety of tumor cells. Normal precursor cells and malignant tumor cells migrate and proliferate, but there is evidence that cells may not be able to engage in both activities at the same time. However, NG2 is involved in promoting both proliferation and motility. The extracellular domain of NG2 sequesters growth factors and binds to both growth factor receptors and extracellular matrix ligands such as fibronectin, collagens and laminin. The cytoplasmic domain is involved in activating Rac, Cdc42 and p130 Cas. PKCα phosphorylates NG2 at Thr2256, triggering the redistribution of NG2 from apical microprocesses to lamellipodia accompanied by enhanced cell motility. ERK phosphorylates NG2 at Thr2314, stimulating cell proliferation. NG2, also known as CSPG4, MCSP, and AN2, is a 400-500 kDa transmembrane chondroitin sulfate proteoglycan (CSPG) with a protein core of approximately 300 kDa. The extracellular region can be proteolytically shed from the cell surface. Mature human NG2 consists of a 2195 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 77 aa cytoplasmic domain. Within aa 1583-2224, human NG2/CSPG4 shares 83% aa sequence identity with mouse and rat CSPG4. NG2 binds to the extracellular matrix proteins Laminin, Tenascin, and Collagens II, V, and VI as well as to the growth factors FGF-2 and PDGF-AA. NG2 is expressed on glial cell progenitors known as O2A cells or NG2 glia. These cells are neuronally responsive and differentiate primarily into oligodendrocytes but also into astrocytes. NG2 associates with PDGF R alpha and the AMPA R subunit GluR2. It is up-regulated on microglial cells during inflammation and contributes to the induction of inflammatory mediators. Various CSPGs in the brain inhibit neurite outgrowth through interactions with Nogo Receptor/NgR1 and NgR3. This recombinant protein product corresponds to the last 5 CSPG repeats, a region which can independently inhibit neurite outgrowth. NG2 is also expressed on vascular mural cells and capillaries. It promotes vascular endothelial cell (EC) migration and angiogenesis through interactions with Galectin-3 and Integrin alpha 3 beta 1 on EC, Plasminogen, and Angiostatin. NG2 is also expressed on a variety of tumors where it contributes to tumor cell adhesion, motility, and invasion. |
包裝清單:
產品編號 | 產品名稱 | 包裝 |
AF5144 | CSPG4/NG2/MCSP Rabbit Polyclonal Antibody (KO Validated) | 50μl |
AZ050 | Western一抗稀釋液 | 50ml |
— | 說明書 | 1份 |
保存條件:
CSPG4/NG2/MCSP 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。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。
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 CSPG4 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 CSPG4 knockout (KO) HeLa cells, using CSPG4 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. |