Phosphotyrosine monoclonal antibody, clone 13F9
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More Files
- More Functions
Western Blot (Cell lysate)
Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is shown to detect by immunoblot (Lane 1) phosphorylated EGFR in an A-431 cell lysate (10 ug per lane) after stimulation with EGF.
The binding of EGF to EGFR causes rapid activation of intrinsic autophosphorylation of multiple tyrosine residues in the cytoplasmic domain of the protein.
This propagates the ERK1/2 signal pathway which regulates cell growth, survival, proliferation and differentiation.
Lane 2 shows total protein in the lysate after coommassie staining.
Minimal reactivity occurs when the antibody is used to stain a lysate from unstimulated A-431 cells.
A 4-20% gradient gel was used for separation prior to transfer to nitrocellulose.
A 1 : 1,000 dilution of Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is used at room temperature for 60 min followed by detection using IRDye™800 Conjugated Goat-a-Mouse IgG [H&L] diluted 1 : 2,500 for 30' at room temperature.
Image was processed and captured using the Odyssey® Infrared Imaging System developed by LI-COR.
IRDye is a trademark of LI-COR, Inc.
Enzyme-linked Immunoabsorbent Assay
ELISA results of PhosphoTyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) tested against BSA conjugates of pT, pY and pS.
Each well was coated with 0.1 ug of conjugate.
The starting dilution of antibody was 1 : 1000 and each point on the X-axis represents a 2-fold dilution.
HRP conjugated Goat-anti-Mouse IgG H&L and TMB substrate were used for detection.
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Specification
Product Description
Mouse monoclonal antibody raised against Phosphotyrosine.
Immunogen
Phosphotyrosine conjugated with KLH.
Host
Mouse
Specificity
Reactivity is specific to phosphotyrosine and minimal cross reactivity is observed against phosphoserine or phosphothreonine.
Form
Liquid
Isotype
IgG1, kappa
Quality Control Testing
Antibody Reactive Against PhosphoTyrosine.
Recommend Usage
Competitive ELISA (1:2000-1:10000)
Western Blot (1:1000)
The optimal working dilution should be determined by the end user.Storage Buffer
In ascites (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is shown to detect by immunoblot (Lane 1) phosphorylated EGFR in an A-431 cell lysate (10 ug per lane) after stimulation with EGF.
The binding of EGF to EGFR causes rapid activation of intrinsic autophosphorylation of multiple tyrosine residues in the cytoplasmic domain of the protein.
This propagates the ERK1/2 signal pathway which regulates cell growth, survival, proliferation and differentiation.
Lane 2 shows total protein in the lysate after coommassie staining.
Minimal reactivity occurs when the antibody is used to stain a lysate from unstimulated A-431 cells.
A 4-20% gradient gel was used for separation prior to transfer to nitrocellulose.
A 1 : 1,000 dilution of Phospho Tyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) is used at room temperature for 60 min followed by detection using IRDye™800 Conjugated Goat-a-Mouse IgG [H&L] diluted 1 : 2,500 for 30' at room temperature.
Image was processed and captured using the Odyssey® Infrared Imaging System developed by LI-COR.
IRDye is a trademark of LI-COR, Inc.Enzyme-linked Immunoabsorbent Assay
ELISA results of PhosphoTyrosine monoclonal antibody, clone 13F9 (Cat # MAB2002) tested against BSA conjugates of pT, pY and pS.
Each well was coated with 0.1 ug of conjugate.
The starting dilution of antibody was 1 : 1000 and each point on the X-axis represents a 2-fold dilution.
HRP conjugated Goat-anti-Mouse IgG H&L and TMB substrate were used for detection. -
Publication Reference
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The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp125FAK and protein kinase C-delta to focal adhesions.
Barry ST, Critchley DR.
Journal of Cell Science 1994 Jul; 107 ( Pt 7):2033.
Application：IF, IP, WB-Ce, Mouse, NIH/3T3 cells.
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Protein-tyrosine kinase activity tightly associated with human type II Fc gamma receptors.
Sarmay G, Pecht I, Gergely J.
PNAS 1994 May; 91(10):4140.
Application：WB-Ce, Human, BL41 cells.
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Immunofluorescent quantification of tyrosine phosphorylation of cellular proteins in whole cells by flow cytometry.
Far DF, Peyron JF, Imbert V, Rossi B.
Cytometry 1994 Apr; 15(4):327.
Application：Flow Cyt, WB-Ce, Human, Jurkat cells.
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Immunohistochemistry of phosphotyrosine residues: identification of distinct intracellular patterns in epithelial and steroidogenic tissues.
Arad-Dann H, Beller U, Haimovitch R, Gavrieli Y, Ben-Sasson SA.
The Journal of Histochemistry and Cytochemistry 1993 Apr; 41(4):513.
Application：IHC-P, Mouse, Rat, Endometrial epithelium lining the uterus.
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The inhibition of EGF-dependent proliferation of keratinocytes by tyrphostin tyrosine kinase blockers.
Dvir A, Milner Y, Chomsky O, Gilon C, Gazit A, Levitzki A.
The Journal of Cell Biology 1991 May; 113(4):857.
Application：IP, RIA, Human, Human keratinocytes.
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The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp125FAK and protein kinase C-delta to focal adhesions.
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