701094

antibody from Invitrogen Antibodies

Targeting: PTK2 FADK, FAK, FAK1, PPP1R71

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

701094

Provider

Invitrogen Antibodies

Product name

FAK Recombinant Rabbit Monoclonal Antibody (5H18L19)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

This antibody is predicted to react with mouse based on sequence homology.

Antibody clone number

5H18L19

Concentration

0.5 mg/mL

References

Submitted references

Opposing effects of in vitro differentiated macrophages sub-type on epithelial wound healing.

Gindele JA, Mang S, Pairet N, Christ I, Gantner F, Schymeinsky J, Lamb DJ
PloS one 2017;12(9):e0184386

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Targeting CXCR4 and FAK reverses doxorubicin resistance and suppresses invasion in non-small cell lung carcinoma.

Dragoj M, Milosevic Z, Bankovic J, Tanic N, Pesic M, Stankovic T
Cellular oncology (Dordrecht) 2017 Feb;40(1):47-62

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FAK in whole cell extracts of Jurkat (lane 1) and K562 (lane 2) cell lines using a FAK recombinant rabbit monoclonal antibody (Product # 701094) at a dilution of 1 µg/mL.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of FAK1 was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with FAK1 Recombinant Rabbit Monoclonal Antibody (Product # 701094) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381). Panel d is a merged image showing membrane and cytoplasmic localization of FAK1. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of FAK1 was done on A549 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª FAK1 Recombinant Rabbit Monoclonal Antibody (701094, red histogram) or with rabbit isotype control (pink histogram) at 2 µg-4 µg/million cells in 2.5% BSA. After incubation at room temperature for 2-3 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig 3 IHC of scratched cells and FN1 regulation/expression. A scratched epithelial cells incubated with different macrophage subtypes after 24h and 72h; DAPI stained in blue, FAK stained in yellow, FN1 stained in red, F-Actin stained in green. B FN1 mRNA regulation in the co-culture cell lysates measured after 24h and 72h Data are displayed as fold regulation compared to control medium containing macrophage maturation factors; Data is expressed as mean +- SD; n = 3 C FN1 expression measured in the supernatants via ELISA after 24h and 72h; Data is expressed as mean +- SD; n = 3 (**** = p>0.0001)

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig 2 IHC of scratched cells and fibronectin expression. A IHC microscopic images of scratched cells at t = 0h, 6h, 24h, 48h and 72h; DAPI in blue, KRT5 in yellow, Ki67 in red. B IHC microscopic images of scratched cells at t = 0h, 6h, 24h, 48h and 72h; DAPI in blue, FAK in yellow, FN1 in red. C ELISA of FN1 in the supernatant after 0h, 6h, 24h, 48h and 72h with and without scratch.