Recombinant
RabMAb

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

Reviews (5)Q&A (1)References (10)

Overview

  • Product name

    Anti-Syndecan-1 antibody [EPR6454]
    See all Syndecan-1 primary antibodies

  • Description

    Rabbit monoclonal [EPR6454] to Syndecan-1

  • 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: ICC/IF, WB, IHC-P, ICC, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Syndecan-1 aa 1-300 (extracellular). The exact sequence is proprietary.
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    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.
  • Immunocytochemistry/ Immunofluorescence - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunocytochemistry/ Immunofluorescence - Anti-Syndecan-1 antibody [EPR6454] (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.

  • Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    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
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



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

  • Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    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
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



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

  • Flow Cytometry - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Flow Cytometry - Anti-Syndecan-1 antibody [EPR6454] (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.
  • Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Western blot - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    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
    All lanes : 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

    Unpurified ab128936 showing positive staining in Normal skin tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

    Unpurified ab128936 showing positive staining in normal tonsil tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

    Unpurified ab128936 showing positive staining in Plasmacytoma tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

    Unpurified ab128936 showing negative staining in Skeletal muscle tissue.

References

This product has been referenced in:

  • Zhu T  et al. Ginsenoside Rg1 attenuates high glucose-induced endothelial barrier dysfunction in human umbilical vein endothelial cells by protecting the endothelial glycocalyx. Exp Ther Med 17:3727-3733 (2019). Read more (PubMed: 30988758) »
  • Indira Chandran V  et al. Ultrasensitive Immunoprofiling of Plasma Extracellular Vesicles Identifies Syndecan-1 as a Potential Tool for Minimally Invasive Diagnosis of Glioma. Clin Cancer Res 25:3115-3127 (2019). Read more (PubMed: 30679164) »
See all 10 Publications for this product

Publishing research using ab128936? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Syndecan-1 antibody [EPR6454]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
50000 cells
Specification
Glioblastoma
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Aug 14 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Syndecan-1 antibody [EPR6454]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
Permeabilization
Yes - 0.05% Tween 20
Specification
Breast
Blocking step
sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Formaldehyde
Read More

Mr. David Ivancic

Verified customer

Submitted Jun 27 2019

Western blot abreview for Anti-Syndecan-1 antibody [EPR6454]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
5 µg
Specification
Retina
Blocking step
Protein Free T20 Blocking Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 31 2018

Flow Cytometry abreview for Anti-Syndecan-1 antibody [EPR6454]

 Excellent
Abreviews
abreview image
Application
Flow Cytometry
Sample
Human Cell (Human myeloma cell line)
Permeabilization
No
Gating Strategy
live cells
Specification
Human myeloma cell line
Read More

Abcam user community

Verified customer

Submitted Jul 11 2016

Immunohistochemistry (Resin sections) abreview for Anti-Syndecan-1 [EPR6454] antibody

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Resin sections)
Sample
Human Tissue sections (Human tonsil)
Specification
Human tonsil
Read More

Abcam user community

Verified customer

Submitted Jan 07 2014

Question

We have recently ordered the anti-syndecan antibody (ab128936) from yourselves, and are currently trying to optimise it for the staining of both human/mouse myeloma cells in paraffin embedded bone sections using both DAB, and immunofluorescence. To date, i have had no success with the staining using concentrations as great as 1/100 (the data sheet suggests 1/500-1/1000), the only thing i am not doing as outlined in the protocol is that i am doing antigen retrieval with proteinase K rather than heat mediated retrieval as suggested. I am going to try the heat retrieval, but our lab has frequently found this unsuitable for bone sections due to the bone lifting from the slide using this method. Any suggestions would be greatly appreciated.

Read More
Answer

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 with some samples. I would recommend using an alternative heat mediated method which may give you better results in this case. This alternative method is to incubate the section in sodium citrate buffer pH9.0 for 30 minutes at 80C. This is best done using a water bath to ensure constant temperature of the buffer.

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