Product: Phospho-HER2/ErbB2 (Tyr1248) Antibody
Catalog: AF3069
Description: Rabbit polyclonal antibody to Phospho-HER2/ErbB2 (Tyr1248)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 185kDa; 138kD(Calculated).
Uniprot: P04626
RRID: AB_2834494

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:500, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-HER2/ErbB2 (Tyr1248) Antibody detects endogenous levels of HER2/ErbB2 only when phosphorylated at Tyrosine 1248.
RRID:
AB_2834494
Cite Format: Affinity Biosciences Cat# AF3069, RRID:AB_2834494.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog; C erb B2/neu protein; CD340; CD340 antigen; Cerb B2/neu protein; CerbB2; Erb b2 receptor tyrosine kinase 2; ERBB2; ERBB2_HUMAN; HER 2; HER 2/NEU; HER2; Herstatin; Human epidermal growth factor receptor 2; Metastatic lymph node gene 19 protein; MLN 19; MLN19; NEU; NEU proto oncogene; Neuro/glioblastoma derived oncogene homolog; Neuroblastoma/glioblastoma derived oncogene homolog; NGL; p185erbB2; Proto-oncogene c-ErbB-2; Proto-oncogene Neu; Receptor tyrosine-protein kinase erbB-2; TKR1; Tyrosine kinase type cell surface receptor HER2; Tyrosine kinase-type cell surface receptor HER2; V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 (neuro/glioblastoma derived oncogene homolog); V erb b2 avian erythroblastic leukemia viral oncogene homolog 2; V erb b2 avian erythroblastic leukemia viral oncoprotein 2; V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian); V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog; Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian);

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P04626 ERBB2_HUMAN:

Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth.

Description:
This gene encodes a member of the epidermal growth factor (EGF) receptor family of receptor tyrosine kinases. This protein has no ligand binding domain of its own and therefore cannot bind growth factors.
Sequence:
MELAALCRWGLLLALLPPGAASTQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTSIISAVVGILLVVVLGVVFGILIKRRQQKIRKYTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLGPASPLDSTFYRSLLEDDDMGDLVDAEEYLVPQQGFFCPDPAPGAGGMVHHRHRSSSTRSGGGDLTLGLEPSEEEAPRSPLAPSEGAGSDVFDGDLGMGAAKGLQSLPTHDPSPLQRYSEDPTVPLPSETDGYVAPLTCSPQPEYVNQPDVRPQPPSPREGPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLTPQGGAAPQPHPPPAFSPAFDNLYYWDQDPPERGAPPSTFKGTPTAENPEYLGLDVPV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Rabbit
100
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P04626 As Substrate

Site PTM Type Enzyme
Phosphorylation
N68 N-Glycosylation
K150 Ubiquitination
K175 Methylation
K175 Ubiquitination
T182 Phosphorylation
S196 Phosphorylation
N259 N-Glycosylation
N530 N-Glycosylation
N571 N-Glycosylation
T686 Phosphorylation P17252 (PRKCA) , P17612 (PRKACA) , Q13237 (PRKG2)
T701 Phosphorylation
S703 Phosphorylation
K716 Ubiquitination
K724 Ubiquitination
S728 Phosphorylation
T733 Phosphorylation
Y735 Phosphorylation
K736 Ubiquitination
K747 Ubiquitination
K753 Ubiquitination
T759 Phosphorylation
Y772 Phosphorylation
S819 Phosphorylation
K854 Ubiquitination
K860 Ubiquitination
T875 Phosphorylation
Y877 Phosphorylation P04626 (ERBB2)
K883 Ubiquitination
K887 Ubiquitination
T900 Phosphorylation Q13131 (PRKAA1)
Y923 Phosphorylation
K937 Ubiquitination
T948 Phosphorylation
S974 Phosphorylation
S977 Phosphorylation
S998 Phosphorylation
S1002 Phosphorylation
T1003 Phosphorylation
Y1005 Phosphorylation
S1007 Phosphorylation
Y1023 Phosphorylation P04626 (ERBB2)
S1049 Phosphorylation
S1050 Phosphorylation
S1051 Phosphorylation
T1052 Phosphorylation
S1054 Phosphorylation
T1060 Phosphorylation
S1066 Phosphorylation
S1073 Phosphorylation
S1078 Phosphorylation
S1083 Phosphorylation
S1100 Phosphorylation
T1103 Phosphorylation
S1107 Phosphorylation
Y1112 Phosphorylation P04626 (ERBB2)
S1113 Phosphorylation
S1122 Phosphorylation
T1124 Phosphorylation
Y1127 Phosphorylation
T1132 Phosphorylation
S1134 Phosphorylation
Y1139 Phosphorylation P04626 (ERBB2)
S1151 Phosphorylation
T1166 Phosphorylation
T1172 Phosphorylation Q9UQM7 (CAMK2A)
S1174 Phosphorylation Q00535 (CDK5)
Y1196 Phosphorylation P04626 (ERBB2)
T1198 Phosphorylation
S1214 Phosphorylation
Y1221 Phosphorylation P04626 (ERBB2)
Y1222 Phosphorylation P04626 (ERBB2)
S1235 Phosphorylation
T1236 Phosphorylation
T1240 Phosphorylation
T1242 Phosphorylation
Y1248 Phosphorylation P00533 (EGFR) , P04626 (ERBB2)

PTMs - P04626 As Enzyme

Substrate Site Source
O43157 (PLXNB1) Y1708 Uniprot
O43157 (PLXNB1) Y1732 Uniprot
P00338 (LDHA) Y10 Uniprot
P00533-1 (EGFR) Y1069 Uniprot
P04626 (ERBB2) Y877 Uniprot
P04626-5 (ERBB2) Y993 Uniprot
P04626 (ERBB2) Y1023 Uniprot
P04626-5 (ERBB2) Y1109 Uniprot
P04626 (ERBB2) Y1112 Uniprot
P04626 (ERBB2) Y1139 Uniprot
P04626-5 (ERBB2) Y1191 Uniprot
P04626-5 (ERBB2) Y1192 Uniprot
P04626 (ERBB2) Y1196 Uniprot
P04626-5 (ERBB2) Y1218 Uniprot
P04626-1 (ERBB2) Y1221 Uniprot
P04626 (ERBB2) Y1222 Uniprot
P04626 (ERBB2) Y1248 Uniprot
P06493 (CDK1) Y15 Uniprot
P35222 (CTNNB1) Y654 Uniprot
Q96N67 (DOCK7) Y1257 Uniprot
Q9BXH1 (BBC3) Y58 Uniprot
Q9BXH1 (BBC3) Y152 Uniprot
Q9BXH1 (BBC3) Y172 Uniprot

Research Backgrounds

Function:

Protein tyrosine kinase that is part of several cell surface receptor complexes, but that apparently needs a coreceptor for ligand binding. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. GP30 is a potential ligand for this receptor. Regulates outgrowth and stabilization of peripheral microtubules (MTs). Upon ERBB2 activation, the MEMO1-RHOA-DIAPH1 signaling pathway elicits the phosphorylation and thus the inhibition of GSK3B at cell membrane. This prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization.

In the nucleus is involved in transcriptional regulation. Associates with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter and activates its transcription. Implicated in transcriptional activation of CDKN1A; the function involves STAT3 and SRC. Involved in the transcription of rRNA genes by RNA Pol I and enhances protein synthesis and cell growth.

PTMs:

Autophosphorylated. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit (Probable). Ligand-binding increases phosphorylation on tyrosine residues. Signaling via SEMA4C promotes phosphorylation at Tyr-1248. Dephosphorylated by PTPN12.

Subcellular Location:

Cell membrane>Single-pass type I membrane protein. Early endosome. Cytoplasm>Perinuclear region. Nucleus.
Note: Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1. Also detected in VPS35-positive endosome-to-TGN retrograde vesicles (PubMed:31138794).

Cytoplasm. Nucleus.

Cytoplasm. Nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth.

Subunit Structure:

Homodimer. Heterodimer with EGFR, ERBB3 and ERBB4. Part of a complex with EGFR and either PIK3C2A or PIK3C2B. May interact with PIK3C2B when phosphorylated on Tyr-1196. Interacts with PLXNB1. Interacts (when phosphorylated on Tyr-1248) with MEMO1. Interacts with MUC1; the interaction is enhanced by heregulin (HRG). Interacts (when phosphorylated on Tyr-1139) with GRB7 (via SH2 domain). Interacts (when phosphorylated on Tyr-1248) with ERBIN. Interacts with KPNB1, RANBP2, EEA1, CRM1 and CLTC. Interacts with PTK6. Interacts with RPA194 and ACTB. Interacts with PRKCABP, SRC and MYOC (By similarity). Interacts (preferentially with the tyrosine phosphorylated form) with CPNE3; this interaction occurs at the cell membrane and is increased in a growth factor heregulin-dependent manner. Interacts with HSP90AA1 and HSP90AB1 in an ATP-dependent manner; the interaction suppresses ERBB2 kinase activity. Interacts with SORL1; this interaction regulates ERBB2 subcellular distribution by promoting its recycling after internalization from endosomes back to the plasma membrane, hence stimulates ERBB2-mediated signaling.

Family&Domains:

Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Adherens junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Calcium signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

References

1). HDAC6 inhibitor TST strengthens the antiproliferative effects of PI3K/mTOR inhibitor BEZ235 in breast cancer cells via suppressing RTK activation. Cell Death & Disease, 2018 (PubMed: 30206202) [IF=9.0]

Application: WB    Species: human    Sample: T47D cells

Fig. 3| HDAC6 inhibitor TST repressed RTK expression. a T47D and b BT474 cells were treated with different concentrations of TST for 24 h.Western blot analyses were performed to detect α-tubulin and its acetylation level. c T47D and d BT474 cells were exposed to increasing doses of TST for 24 h. Cell viability was evaluated by MTT assay. e T47D cells were treated with 10 μM TST and f BT474 cells were treated with 15 μM TST for indicated times. Indicated proteins were detected by western blot

2). Resistance to anti-HER2 therapy associated with the TSC2 nonsynonymous variant c.4349 C > G (p.Pro1450Arg) is reversed by CDK4/6 inhibitor in HER2-positive breast cancer. npj Breast Cancer, 2023 (PubMed: 37160904) [IF=5.9]

Application: WB    Species: Human    Sample: BT474 cell

Fig. 3 Expression of proteins involved in the HER2/AKT/mTOR and cyclinD1/CDK4 signaling pathways in BT474 cell lines treated with lapatinib and/or palbociclib. a Western blot analysis of proteins involved in the HER2/AKT/mTOR signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. b Western blot analysis of proteins involved in the cyclinD1/CDK4 signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. c p-HER2 expression levels under treatment with different drugs in the same cell. d p-HER2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. e p-TSC2 expression levels under treatment with different drugs in the same cell. f p-TSC2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. g p-mTOR expression levels under treatment with different drugs in the same cell. h p-mTOR expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. i p-P70S6K expression levels under treatment with different drugs in the same cell. j p-P70S6K expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. Data are shown as mean ± SD; One-way ANOVA was used to analyze the data in (d, f, h, j). Error bars are SEM.

Application: IHC    Species: Human    Sample: BT474 cell

Fig. 3 Expression of proteins involved in the HER2/AKT/mTOR and cyclinD1/CDK4 signaling pathways in BT474 cell lines treated with lapatinib and/or palbociclib. a Western blot analysis of proteins involved in the HER2/AKT/mTOR signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. b Western blot analysis of proteins involved in the cyclinD1/CDK4 signaling pathway after treatment with lapatinib and/or palbociclib for 48 h. c p-HER2 expression levels under treatment with different drugs in the same cell. d p-HER2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. e p-TSC2 expression levels under treatment with different drugs in the same cell. f p-TSC2 expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. g p-mTOR expression levels under treatment with different drugs in the same cell. h p-mTOR expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. i p-P70S6K expression levels under treatment with different drugs in the same cell. j p-P70S6K expression levels in TSC2-NC, WT, and MT groups treated with the same drugs. Data are shown as mean ± SD; One-way ANOVA was used to analyze the data in (d, f, h, j). Error bars are SEM.

3). Role of EGFR/ErbB2 and PI3K/AKT/e-NOS in Lycium barbarum polysaccharides Ameliorating Endothelial Dysfunction Induced by Oxidative Stress. AMERICAN JOURNAL OF CHINESE MEDICINE, 2019 (PubMed: 31645123) [IF=5.7]

4). Marsdenia tenacissima (Roxb.) Moon injection exerts a potential anti-tumor effect in prostate cancer through inhibiting ErbB2-GSK3β-HIF1α signaling axis. Journal of Ethnopharmacology, 2022 (PubMed: 35595220) [IF=5.4]

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