44-720G

antibody from Invitrogen Antibodies

Targeting: PXN

Provider product page for 44-720G

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [34]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [2]
Other assay [6]

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Product number: 44-720G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Paxillin (Tyr31) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: -20°C

PXN protein structure - 44-720G shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5--figure supplement 1. FAK phosphorylation in Tln Ctr , Tln Ko , Tln Ko+T1V , Kind Ctr , Kind Ko and Kind Ko+K2GFP cells. ( A ) Densitometric quantification of western blot signals of lysates from untreated, EGF- and Mn 2+ -treated Kind Ctr , Tln Ko and Kind Ko cells seeded either on FN or PLL and probed with anti-Tyr-397 phosphorylated FAK (pY397-FAK) antibodies (n=3 independent repeats; significances are calculated with respect to PLL adherent cells; error bars indicate standard error of the mean). ( B , C ) Western blotting of indicated signaling proteins in untreated, EGF- and Mn 2+ -treated Tln Ctr , Tln Ko and Tln Ko cells re-expressing Venus-tagged talin-1 (Tln Ko+T1V ) ( B ), and Kind Ctr , Kind Ko and Kind Ko cells re-expressing GFP-tagged kindlin-2 (Kind Ko+K2GFP ) ( C ) seeded either on FN or PLL. EGF, epidermal growth factor; FAK, focal adhesion kinase; K2GFP, green fluorescent protein-tagged kindlin-2; FN, fibronectin; GFP, green fluorescent protein; PLL, poly-L-lysine; T1V, Venus-tagged full length talin-1. DOI: http://dx.doi.org/

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Focal adhesion association of SRC controls SRC-mediated substrate phosphorylation, focal adhesion formation and cell migration. ( a ) Cell lysates from U2OS cells expressing GFP, SRC-GFP, SRCDeltaSH2-GFP, SRCY527F-GFP or SRCY527FDeltaSH2-GFP were analyzed by Western blotting for pY577-FAK, pY397-FAK, pY118-paxillin, pY31-paxillin, paxillin, GAPDH and GFP. ( b ) Cell lysates from SYF cells expressing GFP, SRC-GFP, SRCDeltaSH2-GFP, SRCY527F-GFP or SRCY527FDeltaSH2-GFP were analyzed by Western blotting for pY577-FAK, pY397-FAK, pY118-paxillin, pY31-paxillin, paxillin, GAPDH and GFP. ( c ) U2OS cells expressing GFP, SRCY527F-GFP or SRCY527FDeltaSH2-GFP (blue) were immunostained to detect F-actin (phalloidin; red) and paxillin (green), and imaged by epi-fluorescence and TIRFM, respectively. Scale bar, 10 mum. ( d ) The number of segmented paxillin-marked FAs within U2OS cells, as described in ( c ). Data are means +- s.e.m. (GFP: n = 11 cells; SRCY527F-GFP: n = 13 cells; SRCY527FDeltaSH2-GFP: n = 12 cells). ** p < 0.01; *** p < 0.001; NS, no significance. ( e ) Size distribution of segmented paxillin-marked FAs of U2OS cells, as described in ( c ). Data are means +- s.e.m. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, no significance. ( f ) The migratory behavior of U2OS cells expressing GFP, SRCY527F-GFP or SRCY527FDeltaSH2-GFP for 6 hrs. Cells were plated for 16 hrs, and then monitored for 6 hrs. (right) These images delineate the trajectory of the GFP-marked cells over a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Focal adhesion kinase activity, but not SRC activity, positively regulates the abundance of SRC_SH2 domain within focal adhesions. ( a ) Total cell lysate from untreated U2OS cells (-) or cells treated with FAKi (focal adhesion kinase inhibitor; 50 muM, 1 h), PP2 (SRC inhibitor; 10 muM, 1 h) or SU6656 (SRC inhibitor; 5 muM, 1 h) were analyzed by Western blotting to detect pY118-paxillin, pY31-paxillin, SRC, FAK, paxillin and GAPDH. ( b ) U2OS cells transfected with pGFP-C1, pGFP-SRC(SH2) or pGFP-SHP2(NSH2) were untreated (-) or treated with FAKi or PP2, and immunostained to detect paxillin. TIRFM images of GFP, GFP-SRC(SH2) or GFP-SHP2(NSH2) (green) and paxillin (red). Bar, 10 mum. The 20 mum x 20 mum areas indicated in the upper images are magnified in the images below. Values indicate Pearson's correlation coefficients (PC) of images of paxillin-Cy5 and GFP, GFP-SRC(SH2) or GFP-SHP2(NSH2) in U2OS cells untreated (-) or treated with FAKi or PP2. Data are means +- s.e.m. (GFP: n = 3 cells (-), n = 4 cells (FAKi), n = 6 cells (PP2); GFP-SRC(SH2): n = 7 cells (-), n = 6 cells (FAKi), n = 7 cells (PP2); GFP-SHP2(NSH2): n = 5 cells (-), n = 5 cells (FAKi), n = 4 cells (PP2)). ** p < 0.01; NS, no significance. ( c ) Time-lapse TIRFM images of U2OS cells co-transfected with mApple-paxillin (red) and pGFP-C1, pGFP-SRC(SH2) or pGFP-SHP2(NSH2) (green) during FAKi or PP2 treatment. The time is shown in hour. Bar, 10 mum. ( d ) The ratio of the average density (intensity p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. Effect of RhoA and RhoB on myosin light chain and paxillin phosphorylation. (A) Pull-down assays of RhoA, RhoB, and RhoC during acute contraction induced with thrombin in HUVECs pretreated with TNF for 8 h. Graphs show active Rho normalized to total Rho levels from three different experiments. (B) TNF-pretreated, siRNA-transfected HUVECs were stimulated with thrombin for the indicated times and immunoblotted for the indicated proteins. (C and D) Quantification of phosphorylation of the indicated proteins from at least three experiments. **, P < 0.005.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Phosphorylated paxillin (Ser-272) is localized in spindle poles during mitosis. ( A ) Mitotic cells were plated on coverslips and were stained with antibodies against phospho-paxillin (pS272-paxillin, pY31-paxillin or pY118-paxillin) and alpha-tublin. Nuclei were stained using DAPI. pS272-paxillin, but not pY31-paxillin and pY118-paxillin, is localized in the spindle poles. Scale bar: 10 mum. Arrows point to spindle poles. ( B ) Z stack images of cells showing connection of microtubule spindle with pS272-paxillin in the poles. Scale bar, 5 mum.