07-5603

antibody from Invitrogen Antibodies

Targeting: NCAM1 CD56, NCAM

Provider product page for 07-5603

Western blot

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [21]
Comments [0]
Validations
Western blot [2]
Immunohistochemistry [1]
Other assay [5]

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Product number: 07-5603 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD56 Monoclonal Antibody (123C3)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 123C3
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20°C

NCAM1 protein structure - 07-5603 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CD56 Monoclonal Antibody (123C3) (Product # 07-5603) and bands in the range of 130-240 kDa corresponding to Neural cell adhesion molecule 1 were observed. Membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), IMR-32 (Lane 2), NK-92 (Lane 3), SK-N-SH (Lane 4), MCF7 (Lane 5), SK-BR-3 (Lane 6) and T-47D (Lane 7) were electrophoresed using 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 (1:1,000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Relative expression was observed between SH-SY5Y, IMR-32, NK-92, SK-N-SH and breast cancer cell lines such as MCF7, SK-BR-3 and T-47D as expected (DOI: 10.1038/s41598-019-45377-8).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CD56 Monoclonal Antibody (123C3) (Product # 07-5603) and a 140 kDa band corresponding to CD56 was observed in SH-SY5Y and SK-N-SH. Membrane enriched cell extracts (12.5 µg lysate) of SH-SY5Y-Untreated (Lane 1), SH-SY5Y-Control (50°C, 3 hrs) (Lane 2), SH-SY5Y-Treated (500 U PNGaseF; 50°C, 3 hr) (Lane 3), SK-N-SH-Untreated (Lane 4), SK-N-SH-Control (50°C, 3 hrs) (Lane 5) and SK-N-SH -Treated (500 U PNGaseF; 50°C, 3 hr) (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000 dilution) using the iBright FL1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). CD56 migrates at ~140 kDa due to multiple glycosylations (Lane 1 and Lane 4). To carry out deglycosylation of the protein, PNGase F Glycan Cleavage Kit was used (Product # A39245). Upon PNGaseF treatment, N-linked glycosylations are cleaved and deglycosylated CD56 can be seen at ~110 kDa (Lane 3 and Lane 6). This mass shift can be attributed to deglycosylation as the control sample

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Immunohistochemistry revealed that the majority of tumor cells were positive CD56 staining in portions (3,3'-diaminobenzidine staining magnification, x500).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Differentiation of neurons after DRG implantation. Panels A-C, fetal DRG, 18 gestational weeks, prior to subcapsular renal implantation in SCID mouse, stained with (A) hematoxylin & eosin; black arrow denotes nerve root, (B) anti-NCAM (green)/anti-synaptophysin (red) antibody, and (C) subtype specific markers anti-RT97 (green)/anti-peripherin (red) antibody. Panels D-E, DRG xenograft, 20 weeks after transplantation, stained with subtype specific markers anti-RT97 (green)/anti-peripherin (red) antibody (D), demonstrates that neuronal differentiation continues in DRG xengrafts in vivo . Peripherin staining (white arrow) identifies nociceptive neurons; cytoplasmic RT97 immunoreactivity (yellow arrow) identifies mechanoreceptive neurons. Cell counting (E) determined the proportion of nociceptive and RT97-immunoreactive mechanoreceptive neurons at 20 weeks after implantation. For each panel, representative images are shown. For assessment of neuronal subtype at least 20 fields were counted, each with a minimum of 50 neurons.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 VZV restriction in mechanoreceptive neurons occurs after viral entry. For each panel, one representative image is shown for staining experiments performed on multiple tissue sections (6-10 sections) for each DRG. (A-C) ORF23 capsid protein is a marker of virion entry as well as the later formation of progeny virions; virion entry is indicated by ORF23 capsid puncta at the nuclear rim (green), co-stained with cellular lamin A/C (A, red), and prior to expression of IE62 (B, red) or IE63 (C, red). (D) Dual staining for ORF23 (green) and cellular PML (red). (E) ORF23 nuclear rim staining was not observed when using pre-immune serum (green); red is N-CAM staining. (F) ORF23 capsid protein is not detected in uninfected DRG, as shown by staining for N-CAM (red) and absence of ORF23 (green). (G-H) Staining of adjacent tissue sections stained with antibody for ORF23 (green) and IE63 (G, red) or RT97 (H, red). Two neurons are circled and shown in both panels. The single 488-channel images for Panel G, shown in greyscale for better visualization, are provided for the circled neurons (1 and 2). The contrast of the G, neuron 2, ORF23 B&W panel, is enhanced so that the discrete ORF23 particles are easier to observe.