Recombinant
RabMAb

Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Overview

  • Product name
    Anti-Syndecan-1 antibody [EPR6454]
    See all Syndecan-1 primary antibodies
  • Description
    Rabbit monoclonal [EPR6454] to Syndecan-1
  • 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: ICC/IF, WB, IHC-P, ICC, 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

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab128936 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
ICC/IF Use a concentration of 2 µg/ml.
WB 1/1000 - 1/10000. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
IHC-P 1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified, use 1/500 - 1/1000.

ICC Use at an assay dependent concentration.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

Images

  • 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.

  • All lanes : Anti-Syndecan-1 antibody [EPR6454] (ab128936) at 1/10000 dilution (purified)

    Lane 1 : Ramos whole cell lysate
    Lane 2 : Raji whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 32 kDa
    Observed band size : 35,100 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Syndecan-1 antibody [EPR6454] (ab128936) at 1/5000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : PC-12 whole cell lysate
    Lane 3 : Mouse heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 32 kDa
    Observed band size : 35,100 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • 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.
  • 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.
  • Lanes 1 - 5 : Anti-Syndecan-1 antibody [EPR6454] (ab128936) at 1/1000 dilution (Unpurified)
    Lane 6 : Anti-Syndecan-1 antibody [EPR6454] (ab128936) at 1/1000 dilution

    Lane 1 : Ramos cell lysate
    Lane 2 : Raji cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : BxPC-3 cell lysate
    Lane 5 : Fetal kidney lysate
    Lane 6 : PC12 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Standard HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 32 kDa
    Observed band size : 35 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 65-100 kDa. We are unsure as to the identity of these extra bands.
  • Unpurified ab128936, at a 1/500 dilution, staining Syndecan in paraffin-embedded Human cervical carcinoma tissue by immunohistochemistry.

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

  • Unpurified ab128936 showing positive staining in Normal skin tissue.

  • Unpurified ab128936 showing positive staining in normal tonsil tissue.

  • Unpurified ab128936 showing positive staining in Plasmacytoma tissue.

  • Unpurified ab128936 showing negative staining in Skeletal muscle tissue.

References

This product has been referenced in:
  • Vähätupa M  et al. Lack of R-Ras Leads to Increased Vascular Permeability in Ischemic Retinopathy. Invest Ophthalmol Vis Sci 57:4898-4909 (2016). IHC-Fr ; Mouse . Read more (PubMed: 27654416) »
  • Ayres Pereira M  et al. Placental Sequestration of Plasmodium falciparum Malaria Parasites Is Mediated by the Interaction Between VAR2CSA and Chondroitin Sulfate A on Syndecan-1. PLoS Pathog 12:e1005831 (2016). IHC . Read more (PubMed: 27556547) »

See all 2 Publications for this product

Customer reviews and Q&As

Application
Flow Cytometry
Sample
Human Cell (Human myeloma cell line)
Permeabilization
No
Gating Strategy
live cells
Specification
Human myeloma cell line
Username

Abcam user community

Verified customer

Submitted Jul 11 2016

Application
Immunohistochemistry (Resin sections)
Sample
Human Tissue sections (Human tonsil)
Specification
Human tonsil
Username

Abcam user community

Verified customer

Submitted Jan 07 2014

I am very sorry to hear of the issues which you have been experiencing with this product. We do recommend use of heat mediated antigen retrieval but do understand that the traditional conditions, e.g. in pressure cooker of microwave, can be an issue wi...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up