Q: I have synovial fluid sample and I want to work on Western bloting can you support me,how can I prepare this sample for WB technique for assay comp(cartilage oligomatrix protein )?

A: We currently sell four antibodies to cartilage oligomeric matrix protein (COMP), all of which are validated for Western blotting. Although we have generated Western blot data using samples from human, bovine and rabbit cartilage, we do not have any testing data derived from synovial fluid samples. You may however find the following publication useful, in which the authors sampled synovial fluid from patients with rheumatoid arthritis and prepared this fluid for Western blot detection of the proteins SP-A and SP-D: PMC 2080374

COMP/Thrombospondin-5 Overexpression Lysate

Novus Biologicals | Catalog # NBP2-09056

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for COMP/Thrombospondin-5 Overexpression Lysate

COMP/Thrombospondin-5 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC211080

Accession#: NM_000095

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

80.9 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.

Type

Overexpression

Scientific Data Images for COMP/Thrombospondin-5 Overexpression Lysate

Western Blot: COMP/Thrombospondin-5 Overexpression Lysate [NBP2-09056]

Western Blot: COMP Overexpression Lysate (Adult Normal) [NBP2-09056] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for COMP.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: COMP/Thrombospondin-5

COMP is a noncollagenous extracellular matrix (ECM) protein. It consists of five identical glycoprotein subunits, each with EGF like and calcium binding (thrombospondin like) domains. Oligomerization results from formation of a five stranded coiled coil and disulfides. Binding to other ECM proteins such as collagen appears to depend on divalent cations. Mutations can cause the osteochondrodysplasias pseudochondroplasia (PSACH) and multiple epiphyseal dysplasia (MED)

Long Name

Cartilage Oligomeric Matrix Protein

Alternate Names

EDM1, EPD1, MED, PSACH, Thrombospondin-5, Thrombospondin5

Gene Symbol

COMP

Additional COMP/Thrombospondin-5 Products

Product Documents for COMP/Thrombospondin-5 Overexpression Lysate

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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 COMP/Thrombospondin-5 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Related Research Areas

Extracellular Matrix Molecules

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