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C3 antibody [M68]

IP/MS
Anti-C3 antibody [M68] used in Immunoprecipitation (IP). GTX02807
Anti-C3 antibody [M68] used in Western Blot (WB). GTX02807
Anti-C3 antibody [M68] used in Western Blot (WB). GTX02807

Cat. No. GTX02807

Host

 Mouse

Clonality

 Monoclonal

Clone Name

 M68

Isotype

 IgG

Application

 WB, IP

Reactivity

 Human
Package
100 μl ($399)
See all C3 products

APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Recommended Dilution
WB 1-5 μg/ml
IP Assay dependent

Note :

WB
For the best detection sensitivity, the samples should be treated under non-boiled and non-reducing conditions.
Not tested in other applications.

Calculated MW

187 kDa. ( Note )

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

Anti-C3 antibody [M68] used in Immunoprecipitation (IP). GTX02807

GTX02807 IP Image

IP/MS

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

Anti-C3 antibody [M68] used in Western Blot (WB). GTX02807

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

Anti-C3 antibody [M68] used in Western Blot (WB). GTX02807

GTX02807 WB Image

IP/MS

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

REFERENCE

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REVIEW

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Package List Price ($)
$ 399