Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23425-68] to Neurocan - BSA and Azide free
  • Suitable for: IHC-P, WB, ICC
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free
    See all Neurocan primary antibodies

  • Description

    Rabbit monoclonal [EPR23425-68] to Neurocan - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, ICCmore details
    Unsuitable for: IHC-Fr or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse brain and hippocampus tissue lysates; Rat brain and hippocampus tissue lysates. IHC-P: Human cerebrum tissue; Mouse cerebrum tissue; Rat cerebrum tissue. ICC: Mouse primary neuron cells.

  • General notes

    ab277536 is the carrier-free version of ab277525. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab277536 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    Constituent: 100% PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR23425-68

  • Isotype

    IgG

  • Research areas

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)

    This data was developed using ab277525, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum (PMID: 9795216). The section was incubated with ab277525 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    All lanes : Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse brain tissue lysate treated with 0.05U chondroitinase ABC for 3h at 37℃,
    Lane 3 : Rat brain tissue lysate
    Lane 4 : Rat brain tissue lysate treated with 0.05U chondroitinase ABC for 3h at 37℃,

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 143 kDa
    Observed band size: 150-300 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab277525, the same antibody clone in a different buffer formulation. 

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

    Neurocan proteolytic cleavage results in two fragments: neurocan-130 (130 kDa) and neurocan-C (150k Da ). Neurocan is a glycoprotein and seen as broad indistinct smears, which on chondroitinase treatment resolve into band of  150 (neurocan-C). The molecular weight observed is consistent with what has been described in the literature (PMID: 10729323).

    Exposure time: 48 seconds.

     

  • Immunocytochemistry - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Immunocytochemistry - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)

    This data was developed using ab277525, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Neurocan with ab277525 at 1/50 (10.6 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain MAP2 at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    -ve control 1: ab277525 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab11267 at 1/200 dilution, followed by ab150077 at 1/1000 dilution.

  • Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    All lanes : Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse hippocampus tissue lysate
    Lane 3 : Mouse skeletal muscle tissue lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 143 kDa
    Observed band size: 130-300 kDa why is the actual band size different from the predicted?



    This data was developed using ab277525, the same antibody clone in a different buffer formulation. 

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

    Neurocan is a glycoprotein. The observed MW is consistent with what has been described in the literature (PMID: 29670169).

    Negative control: mouse skeletal muscle (PMID: 1326557)

    Samples are non-boiled as boiling may cause protein aggregates.

    Exposure time: 26 seconds.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)

    This data was developed using ab277525, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID: 30356641) . The section was incubated with ab277525 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Western blot - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    All lanes : Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat hippocampus tissue lysate
    Lane 3 : Rat skeletal muscle tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 143 kDa
    Observed band size: 130-300 kDa why is the actual band size different from the predicted?



    This data was developed using ab277525, the same antibody clone in a different buffer formulation. 

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

    Neurocan is a glycoprotein. The observed MW is consistent with what has been described in the literature (PMID: 29670169). 

    Negative control: rat skeletal muscle (PMID: 1326557).

    Samples are non-boiled as boiling may cause protein aggregates.

    This blot was developed using a higher sensitivity ECL substrate.

    Exposure time: 103 seconds.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] - BSA and Azide free (ab277536)

    This data was developed using ab277525, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum. The section was incubated with ab277525 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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