PA1-25162

antibody from Invitrogen Antibodies

Targeting: ZYX

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-25162

Provider

Invitrogen Antibodies

Product name

Zyxin Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: HeLa, Rat1.

Concentration

12.7 mg/mL

References

Submitted references

In situ molecular characterization of endoneurial microvessels that form the blood-nerve barrier in normal human adult peripheral nerves.

Ouyang X, Dong C, Ubogu EE
Journal of the peripheral nervous system : JPNS 2019 Jun;24(2):195-206

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HepG2 (Lane 2), COS7 (Lane 3), A549 (Lane 4), A549 serum starved for 15 hr and treated with TGF-β1 (5ng/ml, 24hr) (Lane 5), MCF7 (Lane 6), MCF7 serum starved for 15 hr and treated with Heregulin (25ng/ml for 6hr) (Lane 7), H1975 (Lane 8), Mouse lung (Lane 9) and Mouse colon (Lane 10). The blot was probed with Anti-Zyxin Polyclonal Antibody (Product # PA1-25162, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 75 kDa band corresponding to Zyxin was observed across the cell lines positive for Zyxin (Lanes 1-2), induced in A549 treated with TGF-β1 (Lane 5) and MCF7 treated with Heregulin (Lane 7), while this band was absent in untreated MCF7 (Lane 6). This band was present in Mouse lung (Lane 9) while it was absent in Mouse colon (Lane 10).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Zyxin was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Zyxin Rabbit Polyclonal Antibody (Product # PA1-25162) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization at focal adhesion points. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Endothelial and specialized epithelial-specific proteins. Representative indirect fluorescent digital photomicrographs of cryostat axial sections of normal adult sural nerves show UEA-1-positive endothelial cells (green; A, E, I, M, Q) with expression of CAV1, GJA1, TJP1, ZYX, and CTNNA1 (red; B, F, J, N, R, respectively) restricted to endoneurial microvessels, fenestrated epineurial macrovessels and the perineurium, shown in the merged images at lower and higher magnifications (yellow/orange for endothelial cells). Expression of these proteins by both microvascular and macrovascular endothelial cells and the perineurium suggests non-restrictive barrier, but specialized endothelial and epithelial cell functions in the normal adult human peripheral nerves. White arrows demonstrate positively staining endoneurial microvessels. Blue (4, 6-diamidino-2-phenylindole) staining indicates nuclei. Scale bar 500 mum for A-C, E-G, I-K, M-O, and Q-S and 125 mum for D, H, L, P, and T