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 microl
![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 microl Lane B : Boiled sample, 5 microl Lane C : Boiled and reduced sample, 20 microl](https://www.genetex.com/upload/website/prouct_img/normal/GTX02807/GTX02807_20201130_WB_1_w_23053122_792.webp "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 microl Lane B : Boiled sample, 5 microl Lane C : Boiled and reduced sample, 20 microl")
![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 microl Lane 2 : 5 microl Lane 3 : 2 microl](https://www.genetex.com/upload/website/prouct_img/normal/GTX02807/GTX02807_20201130_WB_w_23053122_846.webp "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 microl Lane 2 : 5 microl Lane 3 : 2 microl")
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 microl
C3 antibody [M68]
GTX02807
ApplicationsImmunoPrecipitation, Western Blot
Product group Antibodies
ReactivityHuman
TargetC3
Overview
- SupplierGeneTex
- Product NameC3 antibody [M68] - IP/MS Validated
- Delivery Days Customer9
- Antibody SpecificityThe 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.
- Application Supplier NoteWB: 1-5 microg/ml. *Optimal dilutions/concentrations should be determined by the researcher.Not tested in other applications.
- ApplicationsImmunoPrecipitation, Western Blot
- CertificationResearch Use Only
- ClonalityMonoclonal
- Clone IDM68
- Concentration1 mg/ml
- ConjugateUnconjugated
- Gene ID718
- Target nameC3
- Target descriptioncomplement C3
- Target synonymsacylation-stimulating protein cleavage product; AHUS5; ARMD9; ASP; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1; C3a; C3a anaphylatoxin; C3b; complement C3; complement component 3; complement component C3a; complement component C3b; CPAMD1; epididymis secretory sperm binding protein Li 62p; HEL-S-62p; prepro-C3
- HostMouse
- IsotypeIgG
- Protein IDP01024
- Protein NameComplement C3
- Scientific DescriptionComplement 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]
- ReactivityHuman
- Storage Instruction-20°C or -80°C,2°C to 8°C
- UNSPSC12352203