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來源
用途
交叉反應性
理論分子量
實際分子量
Rabbit
WB, IP, IF, IHC
H, M, R
44/69/77/134kDa
175kDa
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
1:50-1:100
1:50-1:200
1:50-1:200
1:50-1:200
-
-
-
抗體詳細信息如下::
About this Antibody
Name
EGFR Rabbit Polyclonal Antibody (KO Validated)
Category
Rabbit Polyclonal Antibody (pAb); Primary antibody
Isotype
IgG
Purification method
Affinity purification
Positive samples
-
Cellular location
Cell membrane, Endoplasmic reticulum membrane, Endosome, Endosome membrane, Golgi apparatus membrane, Nucleus membrane, Nucleus, Secreted, Single-pass type I membrane protein
Customer validation
-
About the Immunogen
Immunogen
Recombinant fusion protein of human EGFR (NP_005219.2).
Sequence
QGFFSSPSTSRTPLLSSLSATSNNSTVACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQNPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA
Gene ID
1956
Swiss Prot
P00533
Synonyms
EGFR; ERBB; ERBB1; HER1; NISBD2; PIG61; mENA; epidermal growth factor receptor
Category
Jak/Stat:IL-25 Signaling; Protein Tyrosine Kinase Signaling; Phospholipase Signaling
Background
The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation. Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins. c-Src is involved in phosphorylation of EGFR at Tyr845. The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling. Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation. The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068. A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation. Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation.
包裝清單：
產品編號
產品名稱
包裝
AF5153
EGFR Rabbit Polyclonal Antibody (KO Validated)
50μl
AZ050
Western一抗稀釋液
50ml
—
說明書
1份
保存條件：
EGFR 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 OVCAR3 cells, using EGFR antibody at 1:3000 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 EGFR knockout (KO) HeLa cells, using EGFR antibody at 1:3000 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 prostate using EGFR antibody at dilution of 1:100 (40x lens).
Fig. 4. Immunofluorescence analysis of HeLa cells using EGFR antibody at dilution of 1:100. Blue: DAPI for nuclear staining.
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