Aggrecan Antibody (BC-3) - BSA Free

Novus Biologicals | Catalog # NB600-504

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Canine, Equine, Feline, Guinea Pig, Rabbit, Sheep

Cited:

Human, Mouse, Rat, Bovine, Rabbit

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # BC-3

Format

BSA Free
View all Aggrecan Primary Antibodies »

Product Specifications

Immunogen

ARGSV... synthetic peptide conjugate.

Reactivity Notes

Cross-reacts with Human, Cow, Cat, Dog, Guinea pig, Horse, Pig, Rabbit, Rat and Sheep. Mouse reactivity reported in scientific literature (PMID: 27185069). Not yet tested in other species.

Localization

Secreted

Marker

Chondrocyte Marker

Specificity

Recognizes the aggrecanase (ADAMTS-1, -4 & -5)-generated N-terminal neoepitope ARG after cleavage between amino acids EGE and ARG within the interglobular domain of aggrecanase-catabolised aggrecan.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

250 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 Aggrecan Antibody (BC-3) - BSA Free

Western Blot: Aggrecan Antibody (BC-3)BSA Free [NB600-504]

Western Blot: Aggrecan Antibody (BC-3)BSA Free [NB600-504]

Aggrecan-Antibody-BC-3-Western-Blot-NB600-504-img0002.jpg
Immunocytochemistry/ Immunofluorescence: Aggrecan Antibody (BC-3) - BSA Free [NB600-504]

Immunocytochemistry/ Immunofluorescence: Aggrecan Antibody (BC-3) - BSA Free [NB600-504]

Aggrecan-Antibody-BC-3-Immunocytochemistry-Immunofluorescence-NB600-504-img0001.jpg
Western Blot: Aggrecan Antibody (BC-3)BSA Free [NB600-504]

Western Blot: Aggrecan Antibody (BC-3)BSA Free [NB600-504]

Aggrecan-Antibody-BC-3-Western-Blot-NB600-504-img0003.jpg
Aggrecan Antibody (BC-3) - BSA Free

Western Blot: Aggrecan Antibody (BC-3) - BSA Free [NB600-504] -

Western Blot: Aggrecan Antibody (BC-3) - BSA Free [NB600-504] - Expression levels of related genes during differentiation induction in the 2D environment. (a) qRT-PCR analysis of Sox9, ACAN & collagen II on days 7, 14 & 21 during induction. (b) qRT-PCR analysis of collagen X, Runx2 & annexin V on days 7, 14 & 21 during induction. The results were normalized to B2M mRNA expression. Values are means ± SD (n = 3). (c) Expression of ACAN, collagen II, collagen X & Runx2 was detected by western blotting on day 10 during induction. (d) Expression of ACAN, collagen II, collagen X & Runx2 was detected by western blotting on day 21 during induction. Significant differences from the control group (non-transfection cells) are indicated by *p < 0.05 or **p < 0.01; differences between IHH & SHH transfection groups are indicated by #p < 0.05 or ##p < 0.01 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30858866), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aggrecan Antibody (BC-3) - BSA Free

Western Blot: Aggrecan Antibody (BC-3) - BSA Free [NB600-504] -

Western Blot: Aggrecan Antibody (BC-3) - BSA Free [NB600-504] - Expression levels of related genes during differentiation induction in the RCCS environment. (a) qPCR analysis of Sox9, ACAN & collagen II on days 7, 14 & 21 during induction. (b) qRT-PCR analysis of collagen X, Runx2, & annexin V on days 7, 14 & 21 during induction. The results were normalized to B2M mRNA expression. Values are means ± SD (n = 3). (c) Expression of ACAN, collagen II, collagen X & Runx2 was detected by western blotting on day 10 during differentiation induction. (d) Expression of ACAN, collagen II, collagen X & Runx2 was detected by western blotting on day 21 during differentiation induction. Significant differences from the control group (non-transfection cells) are indicated by *p < 0.05 or ** p < 0.01; differences between IHH & SHH transfection groups are indicated by #p < 0.05 or ##p < 0.01 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30858866), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aggrecan Antibody (BC-3) - BSA Free

Immunocytochemistry/ Immunofluorescence: Aggrecan Antibody (BC-3) - BSA Free [NB600-504] -

P-PRP induces more extracellular matrix-related proteins. (a) Collagen II and aggrecan in the cytoplasm of the coculture cells imaged by fluorescence microscopy. (b) Production of collagen II and aggrecan, as measured by western blot. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30425747), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Aggrecan Antibody (BC-3) - BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:100
Application Notes
This Aggrecan antibody is useful in Immunohistochemistry, ELISA and Western blot. With IHC the antibody works in formalin- or paraformaldehyde-fixed paraffin embedded sections as well as either alcohol-fixed frozen sections or un-fixed snap-frozen sections. In Western blot the antibody detects a variety of epitopes between 50 and 250 kDa. Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. See Little et al. and Caterson et al. 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. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 27185069).

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 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: Aggrecan

The Aggrecan degradation product usually remains within the tissue still complexed to hyaluronan and link protein. Its release from the cartilage usually signals that there has been extensive catabolism of aggrecan, which allows large complexes containing this metabolite to be released from the tissue. Aggrecan degradation products can be found in the synovial fluids of patients with degenerative joint disease.

Alternate Names

ACAN, AGC1, CSPG1, CSPGCP, MSK16, SEDK

Entrez Gene IDs

176 (Human); 58968 (Rat)

Gene Symbol

ACAN

UniProt

P16112

Additional Aggrecan Products

Product Documents for Aggrecan Antibody (BC-3) - BSA Free

Certificate of Analysis

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

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Product Specific Notices for Aggrecan Antibody (BC-3) - 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 Aggrecan Antibody (BC-3) - BSA Free

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Protocols

View specific protocols for Aggrecan Antibody (BC-3) - BSA Free (NB600-504):

This protocol is from the reference Roberts S et al. Matrix metalloproteinases and aggrecanase: their role in disorders of the human intervertebral disc. Spine 25:3005-13 (2000). PubMed: 11145811

1. Immunostaining with enzyme-generated antibodies BC3, 13, 4, and 14 was carried out on 19 discs from patients 11-59 years of age (1 with prolapse, 2 with scoliosis, 2 with spondylolisthesis, and the remainder with degenerative disc disease).

2. Different fixation treatments were used to optimize antigen preservation and staining of the cryosections; for example, 70% ethanol, 100% ethanol, 10% formaldehyde, or no fixation was used.

3. After this treatment, sections to be stained for BC 3 and 14 were digested with keratanase I, II, and chondroitinase ABC for 3 hours, and those to be stained for BC4 and 13 were digested with chondroitinase ABC for only 90 minutes.

4. Labeling was as for paraffin sections with other antibodies.

5. Staining was done on 5 um paraffin sections that then were deparaffinized with xylene and rehydrated through a series of alcohols to phosphate buffered saline (PBS). Staining conditions were optimized for each individual antibody... no pretreatment was found necessary.

6. Sequential blocking of endogenous peroxidase activity was performed with 0.3% hydrogen peroxide in PBS and then 20% normal human and horse serum and 3% bovine serum albumin in PBS.

7. The primary antibody was incubated for 90 minutes at room temperature before labeling with peroxidase linked to a biotin-streptavidin complex using diaminobenzadine as the colorimetric substrate, then washing, dehydrating, and mounting in pertex.

Adjacent control sections were incubated with either a class-matched immunoglobulin raised to an irrelevant antigen or PBS in place of the primary antigen.

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

FAQs for Aggrecan Antibody (BC-3) - BSA Free

Showing  1 - 3 of 3 FAQs Showing All

  • Q: Do you know if this antibody has been used with immunofluorescence?

    A: We have not yet validated this antibody for use in immunofluorescence. In instances such as this, we recommend our Innovators Reward Program. Under the terms and conditions of this program, we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review of this product including an image of either positive or negative staining for testing in a new species and/or application.

  • Q: On behalf of a customer I have a question about antibody NB600-504 anti-ACAN. The note on the datasheet says Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. Do you really mean to say deglycosylated? Are not those reagents actually digesting the aggrecan? Do they also deglycosylate the protein?

    A: Aggrecan is a highly glycosylated protein and this can interfere with the antibody binding to and recognizing the epitope. These enzymes will deglycosylate the protein and expose the epitope for recognition by the antibody

  • Q: Our customer would like to confirm the predicted MW by using NB600-504 to detect Aggrecan. According to info from http://www.uniprot.org/uniprot/P16112, the MW of the target protein may be between 239 to 250 kDa. But this antibody can also be used with ELISA and Western blot, where it detects a variety of epitopes between 50 and 250 kDa is described. May we have the explanation?

    A: There are two isoforms of aggrecan, and this antibody can detect either one or both of them. It depends on which isoforms a particular cell type/tissue/species expresses.

  • Q: Do you know if this antibody has been used with immunofluorescence?

    A: We have not yet validated this antibody for use in immunofluorescence. In instances such as this, we recommend our Innovators Reward Program. Under the terms and conditions of this program, we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review of this product including an image of either positive or negative staining for testing in a new species and/or application.

  • Q: On behalf of a customer I have a question about antibody NB600-504 anti-ACAN. The note on the datasheet says Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. Do you really mean to say deglycosylated? Are not those reagents actually digesting the aggrecan? Do they also deglycosylate the protein?

    A: Aggrecan is a highly glycosylated protein and this can interfere with the antibody binding to and recognizing the epitope. These enzymes will deglycosylate the protein and expose the epitope for recognition by the antibody

  • Q: Our customer would like to confirm the predicted MW by using NB600-504 to detect Aggrecan. According to info from http://www.uniprot.org/uniprot/P16112, the MW of the target protein may be between 239 to 250 kDa. But this antibody can also be used with ELISA and Western blot, where it detects a variety of epitopes between 50 and 250 kDa is described. May we have the explanation?

    A: There are two isoforms of aggrecan, and this antibody can detect either one or both of them. It depends on which isoforms a particular cell type/tissue/species expresses.

  • Q: Do you know if this antibody has been used with immunofluorescence?

    A: We have not yet validated this antibody for use in immunofluorescence. In instances such as this, we recommend our Innovators Reward Program. Under the terms and conditions of this program, we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review of this product including an image of either positive or negative staining for testing in a new species and/or application.

  • Q: On behalf of a customer I have a question about antibody NB600-504 anti-ACAN. The note on the datasheet says Samples must be deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10ug S-GAG of non-deglycosylated aggrecan for optimal epitope recognition. Do you really mean to say deglycosylated? Are not those reagents actually digesting the aggrecan? Do they also deglycosylate the protein?

    A: Aggrecan is a highly glycosylated protein and this can interfere with the antibody binding to and recognizing the epitope. These enzymes will deglycosylate the protein and expose the epitope for recognition by the antibody

  • Q: Our customer would like to confirm the predicted MW by using NB600-504 to detect Aggrecan. According to info from http://www.uniprot.org/uniprot/P16112, the MW of the target protein may be between 239 to 250 kDa. But this antibody can also be used with ELISA and Western blot, where it detects a variety of epitopes between 50 and 250 kDa is described. May we have the explanation?

    A: There are two isoforms of aggrecan, and this antibody can detect either one or both of them. It depends on which isoforms a particular cell type/tissue/species expresses.

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