Anti-Paxillin (phospho Y118) antibody (ab4833)
Key features and details
- Rabbit polyclonal to Paxillin (phospho Y118)
- Suitable for: WB
- Reacts with: Mouse, Human, African green monkey
- Isotype: IgG
Overview
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Product name
Anti-Paxillin (phospho Y118) antibody
See all Paxillin primary antibodies -
Description
Rabbit polyclonal to Paxillin (phospho Y118) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF Recombinant fragmentWB HumanAfrican green monkey
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Immunogen
Synthetic peptide (Human) derived from the region of human paxillin that contains tyrosine 118.
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Positive control
- WB: COS-7, A431, HeLa whole cell lysates
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated paxillin, and ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using a paxillin-derived peptide that is phosphorylated at tyrosine 31. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-Paxillin (phospho Y118) antibody (ab4833)All lanes : Anti-Paxillin (phospho Y118) antibody (ab4833) at 1/500 dilution
Lane 1 : COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate
Lane 2 : COS-7 whole cell lysate treated with 100 uM Pervanadate for 1 hour
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : A431 whole cell lysate treated with 100 uM Pervanadate for 1 hour
Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 6 : HeLa whole cell lysate treated with 100 uM Pervanadate for 1 hour
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/5000 dilution
Predicted band size: 68 kDaA 66 kDa band corresponding to Paxillin pTyr118 was enhanced across treated cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.
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Western blot - Anti-Paxillin (phospho Y118) antibody (ab4833)All lanes : Anti-Paxillin (phospho Y118) antibody (ab4833) at 0.75 µg/ml (3% BSA-TBST buffer)
All lanes : NMµMG (mouse mammary) cells transfected with EGFP-tagged paxillin
Secondary
All lanes : goat F(ab’)2 anti-rabbit IgG alkaline phosphatase
Predicted band size: 68 kDaPeptide Competition: Extracts were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC.
Prior incubation with:
1- no peptide,
2- the non-phosphopeptide corresponding to the immunogen
3- a generic phosphotyrosine containing peptide
4- the phosphopeptide immunogen
Bands were detected using the Tropix WesternStar method.
The data show that only the peptide corresponding to ab4833 blocks the antibody signal, thereby demonstrating the specificity of the antibody.
