Overview

  • Product name
  • Description
    Rabbit polyclonal to Syndecan 4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide surrounding amino acid 184 of human Syndecan 4 (Intracellular domain).

  • Positive control
    • Jurkat cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab24511 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 5 µg/ml. PubMed: 22513363
IHC-P Use at an assay dependent concentration.
IP Use a concentration of 5 - 10 µg/ml. PubMed: 22048770
WB Use a concentration of 1 - 4 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 22 kDa).Can be blocked with Syndecan 4 peptide (ab52121).

Target

  • Function
    Cell surface proteoglycan that bears heparan sulfate.
  • Tissue specificity
    Expressed in epithelial and fibroblastic cells.
  • Sequence similarities
    Belongs to the syndecan proteoglycan family.
  • Post-translational
    modifications
    Shedding is enhanced by a number of factors such as heparanase, thrombin or EGF. Also by stress and wound healing. PMA-mediated shedding is inhibited by TIMP3.
  • Cellular localization
    Membrane. Secreted. Shedding of the ectodomain produces a soluble form.
  • Information by UniProt
  • Database links
  • Alternative names
    • Amphiglycan antibody
    • MGC22217 antibody
    • OTTHUMP00000031788 antibody
    • ryudocan amphiglycan antibody
    • Ryudocan antibody
    • Ryudocan core protein antibody
    • SDC 4 antibody
    • Sdc4 antibody
    • SDC4_HUMAN antibody
    • SYND 4 antibody
    • SYND4 antibody
    • syndecan 4 antibody
    • syndecan 4 (amphiglycan, ryudocan) antibody
    • syndecan proteoglycan 4 antibody
    • Syndecan-4 antibody
    • Syndecan4 antibody
    see all

Images

  • Anti-Syndecan 4 antibody (ab24511) + Jurkat Cell lysate

    Predicted band size: 22 kDa
    Observed band size: 24 kDa
    why is the actual band size different from the predicted?

  • ab24511 (1µg/ml) staining Syndecan 4 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of cellular membrane compartment in the subset of intestinal glands.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab24511 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24511, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemical analysis of mature rat intervertebral disc tissue, staining Syndecan 4 with ab24511.

    Tissue was fixed in 4% paraformaldehyde before incubating with primary antibody (1/200) overnight at 4°C. An AlexaFluor®488-conjugated anti-rabbit IgG (1/200) was used as the secondary antibody.

References

This product has been referenced in:
  • Oikari LE  et al. Cell surface heparan sulfate proteoglycans as novel markers of human neural stem cell fate determination. Stem Cell Res 16:92-104 (2016). ICC/IF ; Human . Read more (PubMed: 26722758) »
  • Li J  et al. Prolyl-4-hydroxylase domain protein 2 controls NF-?B/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus. J Biol Chem 290:7195-207 (2015). Read more (PubMed: 25635047) »
See all 17 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Rat Cell (primary myogenic cells)
Permeabilization
Yes - 0.1% Tween in PBS for 10 min
Specification
primary myogenic cells
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 17 2018

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab104568. The estimated date of delivery is xxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Loading amount
10 µg
Specification
MEF
Gel Running Conditions
Reduced Denaturing (4%-12%)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 05 2009

Question
Answer

Thank you once again for your recent enquiry. I have contacted the originator of this antibody and can confirm it was tested on mouse liver sections in IHC-P. They have provided the testing procedure, which I have copied below. I hope this will be helpful to you. Please do not hesitate to contact us again if you continue to have difficulties with the results from this antibody. Solutions and Reagents: Xylene Ethanol ddH2O Hematotoxylin 10X PBS: 0.58 M Na2HPO4, 0.17 M NaH2PO4, 0.68 M NaCl. Adjust pH to 7.4 10 mM Sodium Citrate Buffer: To prepare 1 liter, add 2.94 g sodium citrate to 1 liter ddH2O. Adjust pH to 6.0. 1% Hydrogen Peroxide: To prepare, add 10 ml 30% H2O2 to 290 ml ddH2O. Blocking Solution: 5% horse serum or goat serum in PBS ABC Reagent Prepare according to manufacturer’s instructions 30 minutes before use. DAB Reagent: Add 6.7 ug of 30% hydrogen peroxide to 10 ml ddH2O; add this mixture to 10 ml of 1 mg/ml DAB (diaminobenzidine tetrahydrochloride) in PBS, filter. Protocol: 1. Deparaffinize/hydrate sections: a) Incubate sections in three washes of xylene for 5 minutes each. b) Incubate sections in two washes of 100% ethanol for 10 minutes each. c) Incubate sections in two washes of 95% ethanol for 10 minutes each. 2. Wash sections twice in ddH2O for 5 minutes each. 3. Wash sections in PBS for 5 minutes. 4. For antigen unmasking, heat sections in 10 mM sodium citrate buffer (pH 6.0) for 1 minute at full power followed by 9 minutes at medium power. (Keep slides fully immersed in buffer and maintain temperature at or just below boiling.) Cool slides for 20 minutes after antigen unmasking. 5. Wash sections in ddH2O three times for 5 minutes each. 6. Incubate sections in 1% hydrogen peroxide for 10 minutes. 7. Wash sections in ddH2O three times for 5 minutes each. 8. Wash section in PBS for 5 minutes. 9. Block each section with 100-400 ul blocking solution for 1 hour at room temperature. 10. Remove blocking solution and add 100-400 ul diluted primary antibody to each section. (Dilute antibody in blocking solution.) Incubate overnight at 4oC. 11. Remove antibody solution and wash sections in PBS three times for 5 minutes each. 12. Add 100-400 ul secondary antibody, diluted in blocking solution, to each section. Incubate 30 minutes at room temperature. 13. If using ABC biotin/avidin method, make ABC reagent according to the manufacturer’s instructions and incubate solution for 30 minutes at room temperature. 14. Remove secondary antibody solution and wash sections three times with PBS for 5 minutes each. 15. Add 100-400 ul ABC reagent to each section and incubate for 30 minutes at room temperature. 16. Remove ABC reagent and wash sections three times in PBS for 5 minutes each. 17. Add 100-400 ul DAB reagent to each section and monitor staining closely. 18. As soon as the section turns brown, immerse slides in ddH2O. 19. If desired, counterstain sections in hematotoxylin for 10 seconds. 20. Wash sections in ddH2O two times for 5 minutes each. 21. Dehydrate sections: a) Incubate sections in 95% ethanol two times for 10 seconds each. b) Repeat in 100% ethanol, incubating sections two times for 10 seconds each. c) Repeat in xylene, incubating sections two times for 10 seconds each. 22. Mount coverslips.

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Question
Answer

Thank you for your enquiry. The antibody has been tested with mouse liver tissue. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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

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