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Purified anti-Phosphothreonine Antibody

Pricing & Availability
Clone
M380A (See other available formats)
Regulatory Status
RUO
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
M380A_PURE_Phosphothreonine_Antibody_1_121720.png
Whole cell lysates (15 μg) from untreated (-) or Calyculin A treated (+) (50 nM for 30 min) HeLa cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with 1.0 μg/mL (1:500 dilution) purified anti-Phosphothreonine antibody (clone M380A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • M380A_PURE_Phosphothreonine_Antibody_1_121720.png
    Whole cell lysates (15 μg) from untreated (-) or Calyculin A treated (+) (50 nM for 30 min) HeLa cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with 1.0 μg/mL (1:500 dilution) purified anti-Phosphothreonine antibody (clone M380A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • M380A_PURE_Phosphothreonine_Antibody_2_121720.png
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.5% Triton-X for 10 minutes, and blocked with 5% FBS for 30 minutes. Cells were then intracellularly stained with a 5.0 µg/mL (1:100 dilution) (panel A) purified anti-Phosphothreonine antibody (clone M380A) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) at 2.5 µg/mL. To demonstrate phospho-specificity of M380A, fixed and permeabilized HeLa cells were pre-treated with lambda protein phosphatase as a control prior to staining with M380A (panel B). Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • M380A_PURE_Phosphothreonine_Antibody_3_121720
    Whole cell extracts (250 µg total protein) prepared from HeLa cells untreated (-) or treated (+) with Calyculin A (50 nM, 30 min) were immunoprecipitated overnight with 2.5 µg of purified mouse IgG1, κ isotype ctrl antibody (Cat. No. 401401) or purified anti-Phosphothreonine antibody (clone M380A). The resulting IP fractions and whole cell extract input (10%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane and probed with purified anti-Phosphothreonine antibody (clone M380A). Lane M: Molecular weight marker.
  • M380A_PURE_Phosphothreonine_Antibody_4_062724
    PBMCs treated with (positive control, filled histogram) or without (negative control, open histogram) Cell Activation Cocktail (without Brefeldin A) (Cat No. 423301) were fixed and permeabilized using True Phos™ Perm Buffer (Cat No. 425401) and intracellularly stained with Purified anti-Phosphothreonine (clone M380A) or Purified anti-Mouse IgG1, κ isotype control (Cat No. 400101) (open histogram, dashed line) (representative for untreated and treated PBMCs), followed by Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322)
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943001 25 µg 95€
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943002 100 µg 235€
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Description

Phosphorylation of specific threonine residues is a post-translational modification event critical for regulating the activity of many proteins.  Protein phosphorylation events can alter protein function through a variety of molecular mechanisms, ranging from conformational changes that can stimulate or inhibit protein function to facilitating protein-protein interactions with binding partners. Antibodies that can detect phosphothreonine residues are valuable tools for identifying novel phospho-sites and characterizing changes in the post-translational state of a broad range of phosphorylated proteins.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide containing phosphorylated threonine
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. For flow cytometric staining using our True-Phos™ Perm Buffer, the suggested use of this reagent is ≤ 1.0 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

RRID
AB_2890864 (BioLegend Cat. No. 943001)
AB_2890864 (BioLegend Cat. No. 943002)

Antigen Details

Function
Cell signaling
Antigen References

1. Zhao et. al. 2016. Sci. Reports. 6:34817.
2. De Verdier C. H. 1952. Nature. 170:804-805.
3. Steinfield J. et. al. 2014. ACS Chem. Biol. 9(5):1104-1112.
4. Mora N. et. al. 2009. Int. Journ. of Peptide and Protein Research. 45(1):53-63.

Gene ID
NA
UniProt
View information about Phosphothreonine on UniProt.org
Go To Top Version: 1    Revision Date: 12.17.2020

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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