MA5-13150

antibody from Invitrogen Antibodies

Targeting: KRT76 HUMCYT2A, KRT2B, KRT2P

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-13150

Provider

Invitrogen Antibodies

Product name

Cytokeratin Pan Type II Monoclonal Antibody (AE3)

Antibody type

Monoclonal

Antigen

Other

Description

MA5-13150 targets Cytokeratin High Molecular Weight in IF and IHC (P) applications and shows reactivity with Bovine, Chicken, Human, mouse, Non-human primate, Rabbit, and Rat samples. The MA5-13150 immunogen is human epidermal Keratin. This antibody detects basic (Type II) cytokeratins: Cytokeratin 1, Cytokeratin 2, Cytokeratin 3, Cytokeratin 4, Cytokeratin 5, Cytokeratin 6, Cytokeratin 7 and Cytokeratin 8.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

AE3

Vial size

500 µL

Concentration

0.2 mg/mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts of T- 47D (Lane 1), PC-12 (Lane 2) and MCF7 (Lane 3). The blot was probed with anti-Cytokeratin Pan Type II Mouse Monoclonal Antibody (Product # MA5-13150, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). Bands between ~ 40-70 kDa corresponding to Cytokeratin Pan Type II were observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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® 2 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).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Formalin-fixed, paraffin-embedded human skin stained with Keratin, HMW antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of epithelial cells.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Cytokeratin Pan Type II was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Cytokeratin Pan Type II Mouse Monoclonal Antibody (MA5-13150, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) 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.