- PA5-13305 - Invitrogen Antibodies CD14 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CD14 in various lysates. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:2,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: Human lung tissue lysate; Lane 2: Human liver tissue lysate; Lane 3: A549 whole cell lysate. Predicted band size: 40 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CD14 in various lysates. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:1,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: Human lung tissue lysate; Lane 2: Human liver tissue lysate. Predicted band size: 40 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CD14 in various lysates. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:1,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: THP-1 whole cell lysate; Lane 2: Human lung tissue lysate; Lane 3: Human liver tissue lysate; Lane 4: A549 whole cell lysate. Predicted band size: 40 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CD14 in various lysates. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:2,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: Human liver lysate; Lane 2: Human lung lysate. Predicted band size: 40 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of PBMC (Lane 1), HL-60 (Lane 2), A549 (Lane 3) and Jurkat (Lane 4). The blot was probed with Anti-CD14 Polyclonal Antibody (Product # PA5-13305, 1:2,000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4,000 dilution). A 50 kDa band corresponding to CD14 was detected across the cell lines tested except for Jurkat which is reported negative for CD14 expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of A549 cells using a CD14 polyclonal antibody (Product # PA5-13305) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CD14 was performed using 70% confluent log phase RAW 264.7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD14 Polyclonal Antibody (Product # PA5-13305) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of CD14 in A549 cells. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:25 followed by Dylight® 488-conjugated goat anti-Rabbit IgG at a dilution of 1:200 (green). Cells were 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized. Immunofluorescence image showing cytoplasm and membrane staining on A549 cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1:500 dilution (red). The nuclear counter stain is DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CD14 was performed using 70% confluent log phase RAW 264.7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD14 Polyclonal Antibody (Product # PA5-13305) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of CD14 in paraffin-embedded human liver tissue. Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:1,000 for 1 hours at room temperature followed by an undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of A549 cells using a CD14 polyclonal antibody (Product # PA5-13305) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of (overlay histogram) of CD14 in Jurkat cells (green line). Samples were incubated with CD14 polyclonal antibody (Product # PA5-13305) using a dilution of 1:25 dilution for 60 min at 37°C followed by Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1:200 dilution for 40 min at 37°C. The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the primary antibody. Isotype control antibody (blue line) was rabbit IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

PA5-13305