Recombinant
RabMAb

Recombinant Anti-Syndecan-1 antibody [EPR6454] - Low endotoxin, Azide free (ab216458)

Overview

  • Product name

    Anti-Syndecan-1 antibody [EPR6454] - Low endotoxin, Azide free
    See all Syndecan-1 primary antibodies
  • Description

    Rabbit monoclonal [EPR6454] to Syndecan-1 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Specificity

    Based on negative staining of mouse and rat spleen and stomach tissues in IHC-P we believe this antibody is unsuitable for IHC-P with mouse and rat. We believe the antibody is suitable for Western blot with mouse and rat based on data from brain, heart, kidney, and spleen lysates. Please contact our Scientific Support team for more information.
  • Tested applications

    Suitable for: IHC-P, ICC, WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Syndecan-1 aa 1-300 (extracellular).
    Database link: P18827

  • Positive control

    • Human colon and cervical carcinoma tissues; Fetal kidney, HeLa, PC12, Raji, Ramos and BxPC-3 cell lysates.
  • General notes

    ab216458 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab216458 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

Images

  • Immunohistochemical staining of paraffin embedded human colon with purified ab128936 at a working dilution of 1/8000. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Immunocytochemistry analysis of A431 cell line labeling Syndecan-1 with Ab195889 at 2.5 μg/ml (top left). Cells were fixed with 4% paraformaldehyde, permaeabilized with 0.1% tritonX-100. DAPI nuclear stain. ab150077, AlexaFluor® 488 Goat anti-Rabbit secondary antibody was used at 2 μg/ml. Confocal image showing membranous staining on A431 cell line.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Overlay histogram showing Raji cells stained with ab128936 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab128936, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive result in 4% paraformaldehyde (10 min) fixed Raji cells used under the same conditions. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Unpurified ab128936, at a 1/500 dilution, staining Syndecan in paraffin-embedded Human colon tissue by immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Unpurified ab128936 showing positive staining in Normal skin tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Unpurified ab128936 showing positive staining in normal tonsil tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Unpurified ab128936 showing positive staining in Plasmacytoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • Unpurified ab128936 showing negative staining in Skeletal muscle tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128936).

  • This IHC data was generated using the same anti-Syndecan 1 antibody clone, EPR6454, in a different buffer formulation (cat# ab128936).

    Unpurified ab128936, at a 1/500 dilution, staining Syndecan in paraffin-embedded Human cervical carcinoma tissue by immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References

ab216458 has not yet been referenced specifically in any publications.

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