MA5-11777

antibody from Invitrogen Antibodies

Targeting: CALD1 CDM, H-CAD, L-CAD

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-11777

Provider

Invitrogen Antibodies

Product name

Caldesmon Monoclonal Antibody (l-CALD)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

MA5-11777 targets Caldesmon LMW in IF, IHC (P), and WB applications and shows reactivity with Human samples.

Antibody clone number

l-CALD

Concentration

0.2 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of Caldesmon LMW using Caldesmon LMW Monoclonal Antibody (Product # MA5-11777) on Rh30 Cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Caldesmon LMW was performed using tissue extracts (30 µg lysate) of Mouse Brain (Lane 1), Mouse Colon (Lane 2) and membrane enriched extract (30 µg lysate) of HT-29 (Lane 3). The blots were probed with Anti-Caldesmon LMW Mouse Monoclonal Antibody (Product # MA5-11777, 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). A 65 kDa band corresponding to Caldesmon was observed across the tissues and cell line tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), 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 uterus stained with Caldesmon antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of smooth muscles.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Caldesmon LMW was performed using HeLa 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 Caldesmon LMW Mouse Monoclonal Antibody (MA5-11777, 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..