Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸

Order now and get it on Tuesday March 02, 2021

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: ICC/IF, WB, IHC-P, Flow Cyt
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

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
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

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 1/10000 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)

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)

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)

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