MA5-32999

antibody from Invitrogen Antibodies

Targeting: FHIT AP3Aase, FRA3B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-32999

Provider

Invitrogen Antibodies

Product name

FHIT Monoclonal Antibody (26H7)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Human FHIT shares 90% and 87% amino acid (aa) sequence identity with mouse and rat FHIT, respectively. Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL. Positive Control - WB: human COLO-320 whole cell, human SW620 whole cell, human SGC-7901 whole cell. ICC/IF: MCF7 cell. Flow: 293T cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

26H7

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FHIT in Lane 1: human COLO-320 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human SGC-7901 whole cell lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with FHIT monoclonal antibody (Product # MA5-32999) using a 0.5 µg/mL dilution, followed by a biotin conjugated goat anti-mouse IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of FHIT using anti-FHIT antibody (Product # MA5-32999). FHIT was detected in a section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL mouse anti-FHIT antibody (Product # MA5-32999) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of FHIT using anti-FHIT antibody (Product # MA5-32999). FHIT was detected in a section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL mouse anti-FHIT antibody (Product # MA5-32999) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of FHIT in 293T cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with FHIT Monoclonal Antibody (26H7) (Product # MA5-32999) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of FHIT in 293T cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with FHIT Monoclonal Antibody (26H7) (Product # MA5-32999) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.