APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
Application |
Recommended Dilution |
1-5 μg/ml |
Assay dependent |
Note :
WB
For the best detection sensitivity, the samples should be treated under non-boiled and non-reducing conditions.
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Not tested in other applications.
Calculated MW
Positive Control
Serum/milk
Product Note
The antibody detects the 50-kDa complement C3 fragment. The identified sequences include thee segments: a.a. 1321-1600, a.a. 200-440, and a.a. 741-930.
PROPERTIES
Form
Liquid
Buffer
PBS, 50% Glycerol
Preservative
No preservative
Storage
Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
Concentration
1 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
Human native protein
Purification
Purified by protein A
Purity
>90% determined by SDS-PAGE
Conjugation
Unconjugated
Note
For laboratory research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Purchasers shall not, and agree not to enable third parties to, analyze, copy, reverse engineer or otherwise attempt to determine the structure or sequence of the product.
TARGET
Synonyms
complement C3 , AHUS5 , ARMD9 , ASP , C3a , C3b , CPAMD1 , HEL-S-62p
Cellular Localization
Secreted
Background
Complement component C3 plays a central role in the activation of complement system. Its activation is required for both classical and alternative complement activation pathways. The encoded preproprotein is proteolytically processed to generate alpha and beta subunits that form the mature protein, which is then further processed to generate numerous peptide products. The C3a peptide, also known as the C3a anaphylatoxin, modulates inflammation and possesses antimicrobial activity. Mutations in this gene are associated with atypical hemolytic uremic syndrome and age-related macular degeneration in human patients. [provided by RefSeq, Nov 2015]
Database
Research Area
DATA IMAGES
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GTX02807 IP Image
The two groups of complement C3 proteins can be purified by GTX02807 C3 antibody [M68] immunoaffinity chromatography from the partially purified milk fractions. Human milk proteins were loaded onto CM-Sepharose 4B column and proteins, including C3, were eluted by different salt (NaCl) concentration. The slat concentration was smaller than 0.3 N. The CM low salt fractions were collected and loaded to the mAb M68-Sepharose column to purify C3. After washing the immunoaffinity column, the captured proteins were eluted by 0.1 N glycine buffer pH 2.4. The eluates were collected into several fractions, E1, E2, E3, E4, E5 and E6. E1-E4 fractions were analyzed by SDS-PAGE and the eluted proteins were stained by CBB. The protein band 2 and 7 as indicated are sliced out and subjected to MS/MS protein identification (by Prottech Inc.), confirming these bands to be complement C3. The relative abundance of peptides matching to C3 is 98% for band 2 and 96% for band 7. For the best detection sensitivity, the samples should be treated under non-boiled and non-reducing conditions. Lane A : Input (partially purified milk fractions) Lane B : Flow through Lane C : Elution 1 Lane D : Elution 2 Lane E : Elution 3 Lane F : Elution 4 Loading : 20 μl
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GTX02807 WB Image
Complement C3 can be detected in human milk by western blot analysis under non-reducing non-boiled conditions using the GTX02807 C3 antibody [M68] as two groups of protein bands. For the best detection sensitivity, the samples should be treated under non-boiled and non-reducing conditions. Lane 1 : 10 μl Lane 2 : 5 μl Lane 3 : 2 μl
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GTX02807 WB Image
The purified complement C3 can be dissociated into four protein bands under reducing and boiled condition. The epitope recognized by GTX02807 C3 antibody [M68] is on the 50-kDa fragment. The epitope recognized by GTX02807 is significantly destroyed by boiling and reducing treatment and requires much more proteins loaded for the detection. The protein band 1, 3, 4, 5, and 6 as indicated are sliced out and subjected to MS/MS protein identification, confirming these bands to be complement C3. The relative abundance of peptides matching to C3 is 97% for band 1, 75% for band 3, 98% for band 4, 91% for band 5, and 45% for band 6. For the best detection sensitivity, the samples should be treated under non-boiled and non-reducing conditions. Lane A : M68 E2 (contained C3 at highest concentration), 1 μl Lane B : Boiled sample, 5 μl Lane C : Boiled and reduced sample, 20 μl
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REFERENCE
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REVIEW
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