Product Name
Paxillin Recombinant Rabbit Monoclonal Antibody [SY23-02] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Paxillin aa 1-59 / 591.
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP
Target Molecular Weight
Predicted band size: 65 kDa
Positive Control
HeLa cell lysate, A431 cell lysate, PANC-1 cell lysate, RAW264.7 cell lysate, C6 cell lysate, Rat testis tissue lysate, SK-OV-3, HeLa, C6, human kidney tissue, mouse testis tissue, human breast carcinoma tissue, mouse ovary tissue, rat testis tissue.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Paxillin is a focal adhesion phosphoprotein that is localized to the cytoskeleton. Phosphorylation of paxillin has been shown to occur in response to PDGF treatment, v-Src transformation or cross-linking of integrins. FAK (focal adhesion kinase) and PYK2 have been shown to phosphorylate paxillin. FAK phosphorylates paxillin specifically on Tyr 118 in vitro. However, FAK phosphorylation does not seem to be required for the recruitment of paxillin to cell adhesion sites. Paxillin may play a role in signal transduction, regulation of cell morphology and the recruitment of structural and signaling molecules to focal adhesions. It has been shown that the amount of paxillin is reduced in mitotic cells by proteolytic downregulation and that paxillin is alternatively phosphorylated on serine rather than on tyrosine and serine during mitosis.
Background References
1. Izumi D et al. CXCL12/CXCR4 activation by cancer-associated fibroblasts promotes integrin 1 clustering and invasiveness in gastric cancer. Int J Cancer 138:1207-19 (2016).
2. Yariswamy M et al. Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction. J Biol Chem 291:19425-36 (2016).
Sequence Similarity
Belongs to the paxillin family.
Post-translational Modification
Phosphorylated by MAPK1/ERK2 (By similarity). Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. Phosphorylation at Ser-244 by CDK5 reduces its interaction with PTK2/FAK1 in matrix-cell focal adhesions (MCFA) during oligodendrocytes (OLs) differentiation. Phosphorylation at Tyr-31 and Tyr-118 by PTK6 promote the activation of RAC1 via CRK/CrKII, thereby promoting migration and invasion. Phosphorylation at Ser-250 by SLK is required for PXN redistribution and cell motility.
Subcellular Location
Cytoplasm, cytoskeleton, Cell junction, focal adhesion, cell cortex.
Synonyms
FLJ16691 antibody
FLJ23042 antibody
Paired box protein Pax 1 antibody
PAX 1 antibody
PAX1 antibody
PAXI_HUMAN antibody
Paxillin alpha antibody
Paxillin antibody
PXN antibody
PXN protein antibody
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Western blot analysis of Paxillin on different lysates with Rabbit anti-Paxillin antibody (ET1607-22) at 1/5,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: PANC-1 cell lysate
Lane 4: RAW264.7 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 65 kDa
Observed band size: 65 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-22) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Western blot analysis of Paxillin on different lysates with Rabbit anti-Paxillin antibody (ET1607-22) at 1/1,000 dilution.
Lane 1: C6 cell lysate (20 µg/Lane)
Lane 2: Rat testis tissue lysate (40 µg/Lane)
Predicted band size: 65 kDa
Observed band size: 65 kDa
Exposure time: 8 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-22) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of SK-OV-3 cells labeling Paxillin with Rabbit anti-Paxillin antibody (ET1607-22) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Paxillin antibody (ET1607-22) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling Paxillin with Rabbit anti-Paxillin antibody (ET1607-22) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Paxillin antibody (ET1607-22) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C6 cells labeling Paxillin with Rabbit anti-Paxillin antibody (ET1607-22) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Paxillin antibody (ET1607-22) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Paxillin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Paxillin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Paxillin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse ovary tissue using anti-Paxillin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-22, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using ET1607-22, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Paxillin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-22, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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