L1CAM Rabbit Polyclonal Antibody
cat.: 0805-5
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 140 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human L1CAM aa 1,208-1,257 / 1,257.
Positive control: A375 cell lysate, A549 cell lysate, HeLa cell lysate, human brain tissue lysate, rat brain tissue lysate, mouse brain tissue lysate, A375, 293T, Hela, human breast cancer tissue, human kidney tissue, mouse kidney tissue, SHG-44.
Subcellular location: Cell membrane, Cell projection, Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:1,000
1:50-1:200
1:50-1:600
1:50-1:100
Uniprot #: SwissProt: P32004 Human | P11627 Mouse | Q05695 Rat
Alternative names: Antigen identified by monoclonal R1 CAML1 CD171 CD171 antigen HSAS HSAS1 Hyd L1 L1 cell adhesion molecule L1-NCAM L1cam L1CAM_HUMAN MASA MIC5 N CAML1 N-CAM-L1 NCAM-L1 NCAML1 Nerve-growth factor-inducible large external glycoprotein Neural cell adhesion molecule L1 NILE OTTHUMP00000025992 S10 SPG1
Images
0805-5_1.jpg Fig1: Western blot analysis of L1CAM on different lysates with Rabbit anti-L1CAM antibody (0805-5) at 1/1,000 dilution.

Lane 1: A375 cell lysate
Lane 2: A549 cell lysate (low expression)
Lane 3: HeLa cell lysate
Lane 4: Human brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 140 kDa
Observed band size: 250 kDa

Exposure time: 1 minute; ECL: K1802;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (0805-5) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
0805-5_2.jpg Fig2: Western blot analysis of L1CAM on different lysates with Rabbit anti-L1CAM antibody (0805-5) at 1/1,000 dilution.

Lane 1: Rat brain tissue lysate
Lane 2: Mouse brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 140 kDa
Observed band size: 220 kDa

Exposure time: 2 minute;

6% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (0805-5) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
0805-5_3.jpg Fig3: Immunocytochemistry analysis of A375 cells labeling L1CAM with Rabbit anti-L1CAM antibody (0805-5) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-L1CAM antibody (0805-5) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
0805-5_4.jpg Fig4: ICC staining NCAML1 in 293T cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
0805-5_5.jpg Fig5: ICC staining NCAML1 in Hela cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
0805-5_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-NCAML1 antibody. Counter stained with hematoxylin.
0805-5_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-L1CAM antibody (0805-5) at 1/600 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (0805-5) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
0805-5_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-NCAML1 antibody. Counter stained with hematoxylin.
0805-5_9.jpg Fig9: Flow cytometric analysis of SHG-44 cells with NCAML1 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.