Recombinant Anti-Paxillin antibody [Y113] (ab32084)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y113] to Paxillin
- Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Paxillin antibody [Y113]
See all Paxillin primary antibodies -
Description
Rabbit monoclonal [Y113] to Paxillin -
Host species
Rabbit -
Specificity
This antibody recognises Paxillin alpha, beta and gamma isoforms.
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Tested applications
Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Dog -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Flow Cyt (intra): HeLa cells. ICC/IF: HeLa cells IP: HeLa whole cell lysate WB: HeLa, RAW 264.7 and mouse heart lysate. IHC-P: Human cerebrum tissue and human breast carcinoma tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 4.17 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y113 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
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Substrate reagent
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32084 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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WB | (10) |
1/1000. Predicted molecular weight: 68 kDa.
For unpurified use at 1/5000 - 1/10000. |
ICC/IF | (7) |
1/50.
For unpurified use at 1/250. |
Flow Cyt (Intra) |
1/100.
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IP | (1) |
1/20.
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IHC-P | (1) |
1/1200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Predicted molecular weight: 68 kDa. For unpurified use at 1/5000 - 1/10000. |
ICC/IF
1/50. For unpurified use at 1/250. |
Flow Cyt (Intra)
1/100. |
IP
1/20. |
IHC-P
1/1200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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: 5829 Human
- Entrez Gene: 19303 Mouse
- Omim: 602505 Human
- SwissProt: P49023 Human
- SwissProt: Q8VI36 Mouse
- Unigene: 446336 Human
- Unigene: 18714 Mouse
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Alternative names
- FLJ16691 antibody
- FLJ23042 antibody
- Paired box protein Pax 1 antibody
see all
Images
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All lanes : Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : PXN knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Anti-PXN antibody [Y113] (ab32084) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32084 was shown to bind specifically to PXN. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PXN knockout cell line ab261892 (knockout cell lysate ab261701). To generate this image, wild-type and PXN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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All lanes : Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Mouse heart lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 68 kDa -
Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Positive staining on human breast carcinoma tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with purified ab32084 at 1/50 dilution (2.88 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis ofHeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution (10µg/ml) (Red). Cells were fixed with4% paraformaldehydeand permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody.Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Immunohistochemistry analysis of paraffin-embedded human cerebrum tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Positive staining on human cerebrum tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Purified ab32084 at 1/20 dilution (0.7µg) immunoprecipitating Paxillin in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg.
Lane 2 (+): ab32084 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32084 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 48 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (180)
ab32084 has been referenced in 180 publications.
- Seetharaman S et al. Microtubules tune mechanosensitive cell responses. Nat Mater 21:366-377 (2022). PubMed: 34663953
- Yu J et al. Dictamnine, a novel c-Met inhibitor, suppresses the proliferation of lung cancer cells by downregulating the PI3K/AKT/mTOR and MAPK signaling pathways. Biochem Pharmacol 195:114864 (2022). PubMed: 34861243
- Kölln LS et al. Label2label: training a neural network to selectively restore cellular structures in fluorescence microscopy. J Cell Sci 135:N/A (2022). PubMed: 35022745
- Wang W et al. Astragaloside IV promotes the angiogenic capacity of adipose-derived mesenchymal stem cells in a hindlimb ischemia model by FAK phosphorylation via CXCR2. Phytomedicine 96:153908 (2022). PubMed: 35026516
- Søreng K et al. ALFY localizes to early endosomes and cellular protrusions to facilitate directional cell migration. J Cell Sci 135:N/A (2022). PubMed: 35099014