701830

antibody from Invitrogen Antibodies

Targeting: FERMT1 C20orf42, FLJ20116, KIND1, UNC112A, URP1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

701830

Provider

Invitrogen Antibodies

Product name

Kindlin Recombinant Rabbit Monoclonal Antibody (7H7L9)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Equine, Bovine, Pig and Rabbit.

Antibody clone number

7H7L9

Concentration

0.5 mg/mL

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 A-431 (Lane 1), A-431 treated with TGF-beta (20 ng/mL for 15 min) (Lane 2), HeLa (Lane 3), HeLa Treated with TGF-beta (20 ng/mL for 15 min) (Lane 4), A549 (Lane 5), A549 Treated with TGF-beta (20 ng/mL for 15 min) (Lane 6), MDA-MB-231 (Lane 7), MDA-MB-231 Treated with TGF-beta (20 ng/mL for 15 min) (Lane 8). The blots were probed with Anti-Kindlin/FERMT1 Recombinant Rabbit Monoclonal Antibody (Product # 701830, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 77 kDa and 53 kDa bands corresponding to Kindlin/FERMT1 was observed according to the treatment. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence was performed on fixed and permeabilized A-431 cells for detection of Kindlin/FERMT1 using Anti- Kindlin/FERMT1 Recombinant Rabbit Monoclonal Antibody (Product # 701830, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Kindlin/FERMT1 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of Kindlin, Panel e) represents control cells with no primary antibody to assess background.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry analysis of Kindlin/FERMT1 was performed on A-431 cells labeled with ABfinity™ Anti-Kindlin/FERMT1 Recombinant Rabbit Monoclonal Antibody (Product# 701830, 2-4 ug/ 1M cells) or with Rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, (Alexa Fluor® 488 conjugate, Product # A27034, 0.4 ug/ml, 1:2500) as represented by the red and yellow histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-Antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer (4468770).