Cyr61/CCN1 Antibody

Western Blot: Cyr61/CCN1 Antibody [NB100-356] - Total protein from Human liver, spleen and HepG2 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-CYR61 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the low expression level of CYR61 in HepG2 cells.

Immunocytochemistry/Immunofluorescence: Cyr61/CCN1 Antibody [NB100-356] - MCF7 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Cyr61/CCN1 (NB100-356) at 1:200 overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at 1:500. Alpha tubulin was used as a co-stain at 1:1000 and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: Cyr61/CCN1 Antibody [NB100-356] - Human endometrium. Photo courtesy of Dr. rer. nat. Isabella Gashaw, University Duisburg-Essen.

Western Blot: Cyr61/CCN1 Antibody [NB100-356] - Detection of cleaved CYR61 in MDA-MB-231 cell lysate using NB 100-356. Photo courtesy of Dr. Lupu's Lab, Northwestern University. Image by Dr. Ingrid Espinoza.

Immunohistochemistry: Cyr61/CCN1 Antibody [NB100-356] - Human endometrium. Photo courtesy of Dr. rer. nat. Isabella Gashaw, University Duisburg-Essen.

Simple Western: Cyr61/CCN1 Antibody [NB100-356] - Image shows a specific band for CYR61 in 0.5 mg/mL of MCF-7 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rb
• Applications: WB, Simple Western, ICC/IF, IHC, IHC-P, B/N
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Cyr61/CCN1 Antibody Summary

• Immunogen: A synthetic peptide made to the human CYR61 protein sequence (between residues 150-250). [UniProt# O00622]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CCN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:200
  • Simple Western 1:400
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:100
  • Block/Neutralize
• Application Notes: Immunohistochemistry was reported in scientific literature. In Western blot a band is observed at ~37 kDa, representing the processed form of CYR61 protein. IF/ICC (PMID 18202125), Blocking/Neutralizing (PMID 22282654).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 37 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Human Artery Whole Tissue Lysate (Adult Whole Lupus)
  Cyr61/CCN1 Overexpression Lysate
  Cyr61/CCN1 Knockout HeLa Cell Lysate

Reactivity Notes

The immunogen sequence has 88% homology to rat and mouse. Rabbit reactivity reported in scientific literature (PMID: 22401280). Mouse reactivity reported in scientific literature (PMID: 27776183).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Cyr61/CCN1 Antibody

• CCN family member 1
• CCN1
• CCN1IGF-binding protein 10
• Cyr61
• Cysteine-rich angiogenic inducer 61
• cysteine-rich, angiogenic inducer, 61
• GIG1
• GIG1cysteine-rich heparin-binding protein 61
• IBP-10
• IGFBP-10
• IGFBP10cysteine-rich, anigogenic inducer, 61
• Insulin-like growth factor-binding protein 10
• protein CYR61
• Protein GIG1

Background

CYR61 (cysteine-rich angiogenic inducer 61), a heparin-binding growth factor, belongs to CCN family whose members are: cysteine-rich 61 (CYR61/CCN1), connective tIssue growth factor (CTGF/CCN2), nephroblastoma overexpressed (NOV/CCN3), and Wnt-induced secreted proteins-1 (WISP-1/CCN4), -2 (WISP-2/CCN5) and -3 (WISP-3/CCN6). CYR61 localizes intracellularly or associates with extracellular matrix at cell surfaces and its reported nuclear localization is surprising, since the primary structure of CYR61 does not include a classic nuclear localization sequence, however, like NOV, EGF, FGF, PDGF, angiogenin, PTHrP etc which lack afore-mentioned sequence, CYR61's nuclear presence may be correlated with transcriptional regulation, and/or mRNA transport. CYR61 plays a key role in wound healing by up-regulating the expression of several genes involved in angiogenesis, inflammation and matrix remodeling including VEGA-A, VEGA-C, MMP1, MMP3, TIMP1, uPA, PAI-1, integrins alpha-3 and alpha-5. CYR61 down-regulates the expression of alpha-1/alpha-2 subunits of collagen type-1 and promotes cell adhesion/adhesive signaling through integrin alpha-6/beta-1, cell migration through integrin alpha-v/beta-5 and cell proliferation through integrin alpha-v/beta-3. CYR61 has been reported to exert its diverse functions via cross-talk with/activating of Wnt, NFkB, AKT and tyrosine kinase signaling pathways.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Cyr61/CCN1 Antibody (NB100-356)(10)

Publications using NB100-356

• Mugahid D, Kalocsay M, Liu X et al. YAP regulates cell size and growth dynamics via non-cell autonomous mediators Elife Jan 9 2020 [PMID: 31913124] (Human)
• Hellinger JW, HUchel S, Goetz L et al. Inhibition of CYR61-S100A4 Axis Limits Breast Cancer Invasion Front Oncol Oct 23 2019 [PMID: 31709177] (Human)
  
  Details:
  The DyLight 550 conjugated version of this antibody was used, NB100-356R.
• Park MH, Kim AK, Manandhar S et al. CCN1 interlinks integrin and Hippo pathway to autoregulate tip cell activity Elife Aug 20 2019 [PMID: 31429823] (B/N)
• Li Y, Adur MK, Kannan A et al. Progesterone Alleviates Endometriosis via Inhibition of Uterine Cell Proliferation, Inflammation and Angiogenesis in an Immunocompetent Mouse Model. PLoS One 2016 Oct 24 [PMID: 27776183] (Mouse)
• Klein R, Stiller S, Gashaw I. Epidermal growth factor upregulates endometrial CYR61 expression via activation of the JAK2/STAT3 pathway Reprod Fertil Dev. 2012 [PMID: 22401280] (ICC/IF, Rabbit)
• Haseley A, Boone S, Wojton J et al. Extracellular matrix protein CCN1 limits oncolytic efficacy in glioma. Cancer Research. 2012 Mar 15 [PMID: 22282654] (B/N, Human)
• Xie JJ, Xu LY, Wu JY et al. Involvement of CYR61 and CTGF in the fascin-mediated proliferation and invasiveness of esophageal squamous cell carcinomas cells. Am J Pathol. 2010 Feb [PMID: 20056838] (WB, IHC-P, Human)
• Dobroff AS, Wang H, Melnikova VO et al. Silencing cAMP-response element-binding protein (CREB) identifies CYR61 as a tumor suppressor gene in melanoma. J Biol Chem. 2009 Sep [PMID: 19632997] (IHC-P, Human)
• Xie JJ, Xu LY, Xie YM. Roles of ezrin in the growth and invasiveness of esophageal squamous carcinoma cells. Int J Cancer. 2009 Jun [PMID: 19165868] (WB, IHC-P, Human)
• Gashaw I, Stiller S, Boing C, Kimmig R, Winterhager E. Premenstrual Regulation of the Pro-Angiogenic Factor CYR61 in Human Endometrium. Endocrinology;149(5):2261-9. May 1 2008. [PMID: 18202125] (ICC/IF, IHC)

Reviews for Cyr61/CCN1 Antibody (NB100-356) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Other.

reviewed by: Anonymous ICC Other 12/12/2014

Summary

• Application: Immunocytochemistry
• Sample Tested: See PMID 22401280
• Species: Other

Blocking

• Blocking Details: See PMID 22401280

Details

• Detection Notes: See PMID 22401280

Comments

• Comments: Published in PMID: 22401280

reviewed by: chandra kurapaty WB Human 06/18/2014

Summary

• Application: Western Blot
• Sample Tested: MDA MB 231 whole cell lysates and conditioned medium
• Species: Human
• Lot: 071103

Blocking

• Blocking Details: 5% milk in TBST 1h RT or 4C O/N

Primary Anitbody

• Dilution Ratio: 1:1000 in 5% milk in TBST, 3h RT or 4C O/N

Secondary Antibody

• Secondary Description: Anti-rabbit IgG, HRP-linked Antibody
• Secondary Manufacturer Cat#: cell signalling 7074
• Secondary Concentration: 1:5000

Details

• Detection Notes: ECL

FAQs for Cyr61/CCN1 Antibody (NB100-356). (Showing 1 - 3 of 3 FAQs).

1. What does the following sentence in the datasheet mean? "Blocking/Neutralizing 50-200 molar excess"
   • NB100-356 has been validated for blocking/neutralizing experiment and in order to do this experiment you need to have the molar concentration of the actual protein in the sample and we recommend using 50-200 molar excess (more than the protein). You can calculate how much of the antibody needs to be added for neutralizing experiment considering NB100-356 is a rabbit IgG and the molecular weight is approximately 160kDa = 160,000 g/mol and based on the molar concentration of the protein, add 50-200 times the amount of the antibody to the samples.
2. I am looking for an antibody that detects human Cyr61 but does not react with mouse Cyr61. Do you have any data to suggest that antibody NB100-356 might work for my experiments?
   • For product NB100-356, we have unfortunately not tested for cross reactivity with the mouse protein. We therefore, would not be able to guarantee that the antibody would not detect the mouse protein.
3. I'd like to add Cyr61/CCN1 Antibody (NB100-356) to liquid culture medium of mouse ES cells. Would this product be suitable in blocking/neutralizing natively secreted Cyr61?
   • Blocking/neutralizing was cited for this antibody in this publication: PMID 22282654. We recommend checking this paper and seeing if their usage is suitable for your intended experiment too.

Bioinformatics

• Gene Symbol: CCN1
• Entrez:
  • 3491(Human)
• Uniprot:
  • O00622(Human)