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
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
1:50-1:100
1:50-1:200
1:50-1:200
1:50-1:200
-
-
-
抗体详细信息如下::
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.
包装清单：
产品编号
产品名称
包装
AF5153
EGFR Rabbit Polyclonal Antibody (KO Validated)
50μl
AZ050
Western一抗稀释液
50ml
—
说明书
1份
保存条件：
EGFR 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。
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片：
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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