MA5-29144

antibody from Invitrogen Antibodies

Targeting: CEACAM6 CD66c, NCA

ELISA

Antibody data

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Validation data

Product number: MA5-29144 - Provider product page
Provider: Invitrogen Antibodies
Product name: CEACAM6 Recombinant Rabbit Monoclonal Antibody (408)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: Applications Reported: This A1exF5 antibody has been reported for use in flow cytometric analysis.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Antibody clone number: 408
Vial size: 100 µL
Concentration: 1.0 mg/mL
Storage: Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

CEACAM6 protein structure - MA5-29144 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CEACAM6 in Lane A: Jurkat Whole Cell Lysate (30 µg). Samples were probed using a CEACAM6 Monoclonal Antibody (Product # MA5-29144) at a 1:500 dilution, followed by a Goat Anti-Rabbit IgG (H+L), Dylight 800 Secondary Antibody at a 1:10000 dilution. Western blot was performed under reducing conditions. Predicted band size:37 kDa. Observed band size:52 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Carcinoembryonic antigen-related cell adhesion molecule 6 was achieved by transfecting A549 with Carcinoembryonic antigen-related cell adhesion molecule 6 specific siRNAs (Silencer® select Product # S9283, S9285). Western blot analysis (Fig. a) was performed using whole cell extracts from the Carcinoembryonic antigen-related cell adhesion molecule 6 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CEACAM6 Recombinant Rabbit Monoclonal Antibody (408) (Product # MA5-29144, 1:1500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Carcinoembryonic antigen-related cell adhesion molecule 6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CEACAM6 Recombinant Rabbit Monoclonal Antibody (408) (Product # MA5-29144) and a 80 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 6 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), MCF7 (Lane 2), MIA PaCa-2 (Lane 3), Hep G2 (Lane 4) and THP-1 (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of Human CEACAM6 in SKBR3 cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with CEACAM6 Recombinant Rabbit Monoclonal Antibody (408) (Product # MA5-29144, 1:300) at 4°C overnight. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Carcinoembryonic antigen-related cell adhesion molecule 6 was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CEACAM6 Recombinant Rabbit Monoclonal Antibody (408) (Product # MA5-29144) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents Hep G2 cells having no expression of CEACAM6. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60 magnification.