Western Blot (WB)
(Figure 1. Western blot analysis of SPARC using anti-SPARC antibody.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat lung tissue lysates,Lane 2: mouse lung tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SPARC antigen affinity purified polyclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for SPARC at approximately 37KD. The expected band size for SPARC is at 35KD.)
Rabbit SPARC Polyclonal Antibody | anti-SPARC antibody
Anti-SPARC Antibody
Gene Names
Sparc; ON; BM-40
Reactivity
Human, Mouse, Rat
Applications
Western Blot, ELISA
Purity
Immunogen Affinity Purified
Synonyms
SPARC; Polyclonal Antibody; Anti-SPARC Antibody; Basement-membrane protein 40; BM-40; Osteonectin; ON; Secreted protein acidic and rich in cysteine; Sparc; Secreted acidic cysteine rich glycoprotein; anti-SPARC antibody
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
IgG
Specificity
No cross reactivity with other proteins.
Purity/Purification
Immunogen Affinity Purified
Form/Format
Lyophilized
Sequence Length
302
Applicable Applications for anti-SPARC antibody
Western Blot (WB), Direct ELISA (EIA)
Application Notes
WB: 0.1-0.5 mug/ml
Direct ELISA: 0.1-0.5 mug/ml
Tested Species: In-house tested species with positive results.
Direct ELISA: 0.1-0.5 mug/ml
Tested Species: In-house tested species with positive results.
Immunogen
E Coli-derived mouse SPARC recombinant protein (Position: H123-L300).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Relevant Detection Systems
It is recommended to use an Enhanced Chemiluminescent Kit with anti-Rabbit IgG (MBS176460) for Western blot.
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time.
Avoid repeated freezing and thawing.
Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of SPARC using anti-SPARC antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat lung tissue lysates,Lane 2: mouse lung tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SPARC antigen affinity purified polyclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for SPARC at approximately 37KD. The expected band size for SPARC is at 35KD.)
Western Blot (WB)
(Figure 1. Western blot analysis of SPARC using anti-SPARC antibody.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat lung tissue lysates,Lane 2: mouse lung tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SPARC antigen affinity purified polyclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for SPARC at approximately 37KD. The expected band size for SPARC is at 35KD.)
Related Product Information for anti-SPARC antibody
Description: Rabbit IgG polyclonal antibody for SPARC detection. Tested with WB, Direct ELISA in Human; Mouse; Rat.
Background: SPARC, secreted protein acidic and rich in cysteine, also known as Osteonectin is a protein that in humans is encoded by the SPARC gene. The human SPARC gene is 26.5 kb long, and contains 10 exons and 9 introns and is located on chromosome 5q31-q33. SPARC is a glycoprotein of 40 kD. SPARC is an acidic, cysteine-rich glycoprotein consisting of a single polypeptide chain that can be broken into 4 domains: 1) an Ca++ binding domains near the glutamic acidic-rich region at the amino terminus (domain I), 2) a cysteine- rich (domain II), 3) a hydrophilic region (domain III) and 4) an EF hand motif at the carboxy terminus region (domain IV). Osteonectin is a glycoprotein in the bone that binds sodium. It is secreted by osteoblasts during bone formation, initiating mineralization and promoting mineral crystal formation. Osteonectin also shows affinity for collagen in addition to bone mineral calcium. A correlation between osteonectin over expression and ampullary cancers and chronic pancreatitis has been found.
Background: SPARC, secreted protein acidic and rich in cysteine, also known as Osteonectin is a protein that in humans is encoded by the SPARC gene. The human SPARC gene is 26.5 kb long, and contains 10 exons and 9 introns and is located on chromosome 5q31-q33. SPARC is a glycoprotein of 40 kD. SPARC is an acidic, cysteine-rich glycoprotein consisting of a single polypeptide chain that can be broken into 4 domains: 1) an Ca++ binding domains near the glutamic acidic-rich region at the amino terminus (domain I), 2) a cysteine- rich (domain II), 3) a hydrophilic region (domain III) and 4) an EF hand motif at the carboxy terminus region (domain IV). Osteonectin is a glycoprotein in the bone that binds sodium. It is secreted by osteoblasts during bone formation, initiating mineralization and promoting mineral crystal formation. Osteonectin also shows affinity for collagen in addition to bone mineral calcium. A correlation between osteonectin over expression and ampullary cancers and chronic pancreatitis has been found.
References
1. Gilmour, D. T., Lyon, G. J., Carlton, M. B. L., Sanes, J. R., Cunningham, J. M., Anderson, J. R., Hogan, B. L. M., Evans, M. J., Colledge, W. H.Mice deficient for the secreted glycoprotein SPARC/osteonectin/BM40 develop normally but show severe age-onset cataract formation and disruption of the lens.EMBO J. 17: 1860-1870, 1998. 2. Goldblum, S. E., Ding, X., Funk, S. E., Sage, E. H.SPARC (secreted protein acidic and rich in cysteine) regulates endothelial cell shape and barrier function.Proc. Nat. Acad. Sci. 91: 3448-3452, 1994. 3. Minn, A. J., Gupta, G. P., Siegel, P. M., Bos, P. D., Shu, W., Giri, D. D., Viale, A., Olshen, A. B., Gerald, W. L., Massague, J.Genes that mediate breast cancer metastasis to lung.Nature 436: 518-524, 2005.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
34,450 Da
NCBI Official Full Name
SPARC
NCBI Official Synonym Full Names
secreted acidic cysteine rich glycoprotein
NCBI Official Symbol
Sparc
NCBI Official Synonym Symbols
ON; BM-40
NCBI Protein Information
SPARC
UniProt Protein Name
SPARC
Protein Family
UniProt Gene Name
Sparc
UniProt Synonym Gene Names
BM-40; ON
Uniprot Description
Appears to regulate cell growth through interactions with the extracellular matrix and cytokines. Binds calcium and copper, several types of collagen, albumin, thrombospondin, PDGF and cell membranes. There are two calcium binding sites; an acidic domain that binds 5 to 8 Ca2+ with a low affinity and an EF-hand loop that binds a Ca2+ ion with a high affinity.
Research Articles on SPARC
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Product Notes
The SPARC sparc (Catalog #AAA1751281) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-SPARC Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's SPARC can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Direct ELISA (EIA). WB: 0.1-0.5 mug/ml Direct ELISA: 0.1-0.5 mug/ml Tested Species: In-house tested species with positive results. Researchers should empirically determine the suitability of the SPARC sparc for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "SPARC, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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