Anti-NCAM1 antibody [EPR21827] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining of ganglia (arrows) in rat colon (PMID : 1705171). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining on mouse cerebrum (PMID : 1705171). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt

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Flow Cytometry - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

Flow cytometric analysis of L-929 (mouse connective tissue fibroblast cell line) cell line (left panel) and Neuro-2a (mouse neuroblastoma cell line) cell line (right panel) labeling NCAM1 with ab220360 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells. Negative control : L-929.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling NCAM1 with ab220360 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining of ganglion (arrow) in mouse stomach (PMID : 1705171). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rab it IgG H&L (HRP), Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling NCAM1 with ab220360 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Neuro-2a cell line.

Negative control : L-929 PMID : 9696812).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

• IP

Supplier Data

Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

NCAM1 was immunoprecipitated from 0.35 mg of rat brain lysate with ab220360 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Rat brain lysate 10 μg (Input).

Lane 2 : ab220360 IP in rat brain lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220360 in rat brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

The 120,140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

All lanes:

Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (<a href='/en-us/products/primary-antibodies/ncam1-antibody-epr21827-ab220360'>ab220360</a>)

Predicted band size: 94 kDa

Observed band size: 120-200 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] - BSA and Azide free (AB231826)

NCAM1 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab220360 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Mouse brain lysate 10 μg (Input).

Lane 2 : ab220360 IP in mouse brain lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220360 in mouse brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

The 120,140 and 180kDa bands are different isoforms as reported in the literature (PMID : 26288071).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220360).

All lanes:

Immunoprecipitation - Anti-NCAM1 antibody [EPR21827] (<a href='/en-us/products/primary-antibodies/ncam1-antibody-epr21827-ab220360'>ab220360</a>)

Predicted band size: 94 kDa

Observed band size: 120-200 kDa

false