MA5-14050

antibody from Invitrogen Antibodies

Targeting: MME CALLA, CD10, NEP

Antibody data

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Validation data

Product number: MA5-14050 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD10 Monoclonal Antibody (56C6)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14050 targets CD10 in WB, IF, FACS and IHC (P) applications and shows reactivity with Human and Rat samples. This antibody is not recommended for LNCAP cells in Western blot applications. The MA5-14050 immunogen is recombinant external domain of CD10 protein.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 56C6
Vial size: 500 µL
Concentration: Conc. Not Determined
Storage: 4° C

MME protein structure - MA5-14050 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of CD10 was performed by loading 25 µg of Daudi (Lane 1) and rat kidney (Lane 2) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a CD10 monoclonal antibody (Product # MA5-14050) at a dilution of 1:100 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at approx. 100 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockout of CD10 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR1056041_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CD10 was performed by loading 30 µg of LNCaP wild type (Lane 1) and LNCaP CD10 KO (Lane 2) membrane extracts. The samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-CD10 Monoclonal Antibody (56C6)(Product # MA5-14050) using 1:500 dilution and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CD10.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CD10 Mouse Monoclonal Antibody (Product # MA5-14050) and a 100 kDa band corresponding to CD10 was observed in cell lines and tissues tested except for PC-3, HeLa, MCF7 cell lines and Mouse liver which are reported to be negative. Membrane enriched extracts (30 µg lysate) of LnCaP (Lane 1), PC-3 (Lane 2), Raji (Lane 3), Ramos (Lane 4), HeLa (Lane 5), MCF7 (Lane 6), Mouse Kidney (Lane 7), Rat Kidney (Lane 8) and Mouse Liver (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:100 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 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: Formalin-fixed, paraffin-embedded human small intestine stained with CD10 antibody using peroxidase-conjugate and DAB chromogen. Note surface staining of epithelial cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD10/MME (green) showing staining in the in the membrane and cytoplasm of human kidney tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a CD10/MME monoclonal antibody (Product # MA5-14050) in 3% BSA-PBS at a dilution of 1:10 and incubated overnight at 4ºC in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD10/MME (green) showing staining in the in the membrane and cytoplasm of human small intestine tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a CD10/MME monoclonal antibody (Product # MA5-14050) in 3% BSA-PBS at a dilution of 1:10 and incubated overnight at 4ºC in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD10/MME (green) showing staining in the in the membrane and cytoplasm of rat kidney tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a CD10/MME monoclonal antibody (Product # MA5-14050) in 3% BSA-PBS at a dilution of 1:10 and incubated overnight at 4ºC in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded rat small intestine stained with CD10 antibody using peroxidase-conjugate and DAB chromogen. Note surface staining of epithelial cells.