Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rabbit

Cited:

Human, Mouse, Rabbit

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Block/Neutralize, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Western Blot, Block/Neutralize, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Functional Assay, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Cyr61/CCN1 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to the human CYR61 protein sequence (between residues 150-250). [UniProt# O00622]

Reactivity Notes

Rabbit reactivity reported in scientific literature (PMID: 22401280). Mouse reactivity reported in scientific literature (PMID: 27776183).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for Cyr61/CCN1 Antibody - BSA Free

Immunohistochemistry: Cyr61/CCN1 Antibody - BSA Free [NB100-356]

Immunohistochemistry: Cyr61/CCN1 Antibody - BSA Free [NB100-356]

Cyr61-CCN1-Antibody-Immunohistochemistry-NB100-356-img0007.jpg
Western Blot: Cyr61/CCN1 AntibodyBSA Free [NB100-356]

Western Blot: Cyr61/CCN1 AntibodyBSA Free [NB100-356]

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 - BSA Free [NB100-356]

Immunocytochemistry/ Immunofluorescence: Cyr61/CCN1 Antibody - BSA Free [NB100-356]

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.
Western Blot: Cyr61/CCN1 AntibodyBSA Free [NB100-356]

Western Blot: Cyr61/CCN1 AntibodyBSA Free [NB100-356]

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 - BSA Free [NB100-356]

Immunohistochemistry: Cyr61/CCN1 Antibody - BSA Free [NB100-356]

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

Immunohistochemistry: Cyr61/CCN1 Antibody - BSA Free [NB100-356]

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

Simple Western: Cyr61/CCN1 AntibodyBSA Free [NB100-356]

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.
Cyr61/CCN1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Cyr61/CCN1 Antibody - BSA Free [NB100-356] -

Immunocytochemistry/ Immunofluorescence: Cyr61/CCN1 Antibody - BSA Free [NB100-356] - P4 inhibits E2-dependent cell proliferation & angiogenesis in ectopic lesions.Sections of the ectopic lesions collected from E2 or E2 plus P4-treated recipients (D16, n = 6) were subjected to histological examination. (A) Representative images (20X) showing H&E & Trichrome staining, or IHC analysis using antibody against myofibroblast biomarker alpha SMA, uterine epithelial biomarker KRT11, or uterine stromal biomarker VIM, respectively. (B) Representative images (20X) showing IHC analysis using antibodies against cell proliferation biomarker KI67, smooth muscle biomarker alpha SMA, endothelial cells CD31, or an angiogenetic regulator CCN1, respectively. The numbers of KI67-positive cells, the perimeters of the supporting blood vessels, & the immunostaining intensities of CCN1 & CD31 were analyzed by ImageJ software. The numerical values were analyzed by One-way ANOVA followed by Dunnett’s post hoc test & expressed as mean ± SEM. Statistical significance is defined as #: p < 0.05, *: p<0.01. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0165347), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Cyr61/CCN1 Antibody - BSA Free

Application
Recommended Usage

Block/Neutralize

reported in scientific literature (PMID 22282654)

Immunocytochemistry/ Immunofluorescence

reported in scientific literature (PMID 18202125)

Immunohistochemistry

reported in scientific literature

Immunohistochemistry-Paraffin

1:100

Simple Western

1:400

Western Blot

1:200
Application Notes

In Western blot a band is observed at ~37 kDa, representing the processed form of CYR61 protein. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in MCF-7 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:400, apparent MW was 39 kDa

Reviewed Applications

Read 2 reviews rated 5 using NB100-356 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Cyr61/CCN1

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.

Long Name

Cysteine-Rich, Angiogenic Inducer 61

Alternate Names

CCN1, GIG1, IGFBP-10

Entrez Gene IDs

3491 (Human)

Gene Symbol

CCN1

UniProt

O00622 (Human)

Additional Cyr61/CCN1 Products

Product Documents for Cyr61/CCN1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Cyr61/CCN1 Antibody - BSA Free

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.

Citations for Cyr61/CCN1 Antibody - BSA Free

Customer Reviews for Cyr61/CCN1 Antibody - BSA Free (2)

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Customer Images

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  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: See PMID 22401280
    Species: Other
    Verified Customer | Posted 12/12/2014
  • Name: chandra kurapaty
    Application: Western Blot
    Sample Tested: MDA MB 231 whole cell lysates and conditioned medium
    Species: Human
    Verified Customer | Posted 06/18/2014
    Cyr61/CCN1 Antibody - BSA Free NB100-356

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Protocols

View specific protocols for Cyr61/CCN1 Antibody - BSA Free (NB100-356):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Cyr61/CCN1 Antibody - BSA Free

Showing  1 - 3 of 3 FAQs Showing All

  • Q: 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?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What does the following sentence in the datasheet mean? "Blocking/Neutralizing 50-200 molar excess"

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What does the following sentence in the datasheet mean? "Blocking/Neutralizing 50-200 molar excess"

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: What does the following sentence in the datasheet mean? "Blocking/Neutralizing 50-200 molar excess"

    A: 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.

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