Anti-Paxillin (phospho S126) antibody (ab24402)
Key features and details
- Rabbit polyclonal to Paxillin (phospho S126)
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Paxillin (phospho S126) antibody
See all Paxillin primary antibodies -
Description
Rabbit polyclonal to Paxillin (phospho S126) -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Chicken -
Immunogen
Synthetic peptide corresponding to Human Paxillin (phospho S126).
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Positive control
- WB: RAW 264.7 cells treated with LPS. A549. HepG2 IHC-P: human breast carcinoma ICC/IF: NIH/3T3
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated paxillin. The final product is generated by affinity chromatography using a paxillin-derived peptide that is phosphorylated at serine 126. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab24402 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
1/20. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/250.
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WB |
1/1000. Detects a band of approximately 68 kDa.
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Notes |
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IHC-P
1/20. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/250. |
WB
1/1000. Detects a band of approximately 68 kDa. |
Target
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Function
Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). -
Sequence similarities
Belongs to the paxillin family.
Contains 4 LIM zinc-binding domains. -
Post-translational
modificationsPhosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. -
Cellular localization
Cytoplasm > cytoskeleton. Cell junction > focal adhesion. - Information by UniProt
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Database links
- Entrez Gene: 395832 Chicken
- Entrez Gene: 5829 Human
- Entrez Gene: 19303 Mouse
- Omim: 602505 Human
- SwissProt: P49024 Chicken
- SwissProt: P49023 Human
- SwissProt: Q8VI36 Mouse
- Unigene: 446336 Human
see all -
Alternative names
- FLJ16691 antibody
- FLJ23042 antibody
- Paired box protein Pax 1 antibody
see all
Images
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All lanes : Anti-Paxillin (phospho S126) antibody (ab24402) at 1/1000 dilution
Lane 1 : Lysate from untreated RAW 264.7 cells
Lane 2 : Lysate from RAW 264.7 cells treated with 1 µg/mL LPS for 60 minutes
Lane 3 : Lysate from RAW 264.7 cells treated with 1 µg/mL LPS for 60 minutes with non-phosphorylated peptide corresponding to the immunogen
Lane 4 : Lysate from RAW 264.7 cells treated with 1 µg/mL LPS for 60 minutes with generic phosphoserine-containing peptide
Lane 5 : Lysate from RAW 264.7 cells treated with 1 µg/mL LPS for 60 minutes with phosphopeptide corresponding to Paxillin [pS91 ]
Lane 6 : Lysate from RAW 264.7 cells treated with 1 µg/mL LPS for 60 minutes with phosphopeptide immunogen
Secondary
All lanes : Goat F(ab)2 anti-rabbit IgG HRP conjugate.
Observed band size: 68 kDa why is the actual band size different from the predicted?
Lysates were resolved on a 10% polyacrylamide gel and transferred to PVDF. The membrane was blocked with a 5% milk-TBST buffer for one hour at room temperature, and then incubated with the Paxillin [pS126] antibody for two hours at room temperature in a 1% milk-TBST buffer, following its prior incubation with or without peptides. Following incubation with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that only the phosphopeptide corresponding to Paxillin [pS 126 ] blocks the antibody signal, thereby demonstrating the specificity of the antibody. No competition was seen following incubation with paxillin phosphopeptides to S130, S178, S380, S455, or S479 (not shown). The antibody was also shown to be specific using 293 cells transfected with wild-type EGFP-tagged human paxillin treated with EGF (not shown). -
All lanes : Anti-Paxillin (phospho S126) antibody (ab24402) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : A549 whole cell lysate treated with 0.1 ug/mL of HGF treatment for 10 minutes
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : HepG2 whole cell lysate with 0.1 ug/mL of HGF treatment for 10 minutes
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/1000 dilution
Observed band size: 68 kDa why is the actual band size different from the predicted?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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Immunohistochemical analysis of human breast carcinoma labeling Paxillin (phospho S126) with ab24402 at 1/20 dilution (right) compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab24402 diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting
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Immunofluorescence analysis of 70% confluent log phase NIH/3T3 cells treated with 0.5 ug of PDGF for 10 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. Labeling Paxillin (phospho S126) with ab24402 at 1/250 dilution in 0.1% BSA and incubated for 3 hours at room temperature. Followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Rhodamine Phalloidin at 1/300 dilution. Panel d is a merged image showing punctuated nuclear localization. Panel e is untreated cell with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab24402 has been referenced in 1 publication.
- Boeckeler K et al. Manipulating signal delivery - plasma-membrane ERK activation in aPKC-dependent migration. J Cell Sci 123:2725-32 (2010). WB ; Rat . PubMed: 20647370