Name: Anti-Syndecan-1 antibody [EPR6454]
Brand: RabMAb
SKU: ab128936
Rating: 5 (6 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR6454] to Syndecan-1
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
Reacts with: Mouse, Rat, Human

Carrier Free PE

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: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Human colon and cervical carcinoma tissues; Fetal kidney, HeLa, PC12, U266B1, Raji, Ramos and BxPC-3 cell lysates.
General notes
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.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR6454
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
Recombinant Protein
- Recombinant Human Syndecan-1 protein (His tag) (ab124600)

The Abpromise guarantee

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
Flow Cyt (Intra)		1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF	(1)	Use a concentration of 2 µg/ml.
WB	(2)	1/1000 - 1/10000. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
IHC-P	(1)	1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. For unpurified, use 1/500 - 1/1000. 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.

Notes
Flow Cyt (Intra) 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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. 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.

Function

Cell surface proteoglycan that bears both heparan sulfate and chondroitin sulfate and that links the cytoskeleton to the interstitial matrix.
Sequence similarities

Belongs to the syndecan proteoglycan family.
Cellular localization

Membrane.
Target information above from: UniProt accession P18827 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6382 Human
- Entrez Gene: 20969 Mouse
- Entrez Gene: 25216 Rat
- Omim: 186355 Human
- SwissProt: P18827 Human
- SwissProt: P18828 Mouse
- SwissProt: P26260 Rat
- Unigene: 224607 Human
- Unigene: 665958 Human
- Unigene: 2580 Mouse
- Unigene: 11176 Rat
see all
Alternative names
- CD_antigen antibody
- CD138 antibody
- CD138 antigen antibody
- heparan sulfate proteoglycan fibroblast growth factor receptor antibody
- SDC antibody
- Sdc1 antibody
- SDC1_HUMAN antibody
- SYND1 antibody
- Syndecan 1 antibody
- Syndecan antibody
- syndecan proteoglycan 1 antibody
- Syndecan-1 antibody
see all

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 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)

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

Intracellular Flow Cytometry analysis of U266B1 (Human multiple myeloma B lymphocyte) cells labeling Syndecan-1 with purified ab128936 at 1/1000 dilution (1µg/mL) (Red). A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Gated on viable cells.
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
Flow Cytometry (Intracellular) - 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)

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)

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.
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.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Unpurified ab128936 showing positive staining in Normal human skin tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Unpurified ab128936 showing positive staining in normal human tonsil tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Unpurified ab128936 showing positive staining in human Plasmacytoma tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Unpurified ab128936 showing negative staining in human Skeletal muscle tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-Syndecan-1 antibody [EPR6454] (ab128936)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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

ab128936 has been referenced in 26 publications.

Zhang Y et al. Endothelial progenitor cells-derived exosomal microRNA-21-5p alleviates sepsis-induced acute kidney injury by inhibiting RUNX1 expression. Cell Death Dis 12:335 (2021). PubMed: 33785732
Zhang Y et al. Phosphatase Shp2 regulates biogenesis of small extracellular vesicles by dephosphorylating Syntenin. J Extracell Vesicles 10:e12078 (2021). PubMed: 33732417
Cerezo-Magaña M et al. Hypoxic Induction of Exosome Uptake through Proteoglycan-Dependent Endocytosis Fuels the Lipid Droplet Phenotype in Glioma. Mol Cancer Res 19:528-540 (2021). PubMed: 33288734
Hu X et al. Syndecan-4 promotes vascular beds formation in tissue engineered liver via thrombospondin 1. Bioengineered 11:1313-1324 (2020). PubMed: 33251971
Huang X et al. Both UFH and NAH alleviate shedding of endothelial glycocalyx and coagulopathy in LPS-induced sepsis. Exp Ther Med 19:913-922 (2020). PubMed: 32010252

View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Kidney H9 Organoid)

Permeabilization

Yes - 0.05% Tween 20

Specification

Kidney H9 Organoid

Blocking step

Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 05 2020

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

Abcam user community

Verified customer

Submitted Aug 14 2019

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

MR. David Ivancic

Verified customer

Submitted Jun 27 2019

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

Abcam user community

Verified customer

Submitted Oct 31 2018

Application

Flow Cytometry

Sample

Human Cell (Human myeloma cell line)

Permeabilization

Gating Strategy

live cells

Specification

Human myeloma cell line

Abcam user community

Verified customer

Submitted Jul 11 2016

Application

Immunohistochemistry (Resin sections)

Sample

Human Tissue sections (Human tonsil)

Specification

Human tonsil

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.

Abcam community

Verified customer

Asked on Feb 20 2013

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.

Abcam Scientific Support

Answered on Feb 20 2013

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (26)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Syndecan-1 antibody [EPR6454]

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

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

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

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

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