Recombinant
RabMAb

Recombinant Anti-C3 antibody [EPR19394] (ab200999)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19394] to C3
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-C3 antibody [EPR19394]
    See all C3 primary antibodies

  • Description

    Rabbit monoclonal [EPR19394] to C3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human C3 aa 1600 to the C-terminus. The exact sequence is proprietary.
    Database link: P01024

  • Positive control

    • WB: Human fetal liver, fetal heart and fetal kidney lysates; Mouse brain, kidney, spleen and heart lysates; Rat brain, spleen, heart and kidney lysates; Human serum, plasma and milk; HepG2 whole cell lysate; Mouse and rat plasma. IHC-P: Mouse liver, cerebral cortex and cardiac muscle tissues; Rat kidney, liver and lung tissues.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab200999 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 187, 115 ,68 ,43 kDa (predicted molecular weight: 187 kDa).

Target

  • Function

    C3 plays a central role in the activation of the complement system. Its processing by C3 convertase is the central reaction in both classical and alternative complement pathways. After activation C3b can bind covalently, via its reactive thioester, to cell surface carbohydrates or immune aggregates.
    Derived from proteolytic degradation of complement C3, C3a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes.

  • Tissue specificity

    Plasma.

  • Involvement in disease

    Defects in C3 are the cause of complement component 3 deficiency (C3D) [MIM:120700]. A rare defect of the complement classical pathway. Patients develop recurrent, severe, pyogenic infections because of ineffective opsonization of pathogens. Some patients may also develop autoimmune disorders, such as arthralgia and vasculitic rashes, lupus-like syndrome and membranoproliferative glomerulonephritis.
    Genetic variation in C3 is associated with susceptibility to age-related macular degeneration type 9 (ARMD9) [MIM:611378]. ARMD is a multifactorial eye disease and the most common cause of irreversible vision loss in the developed world. In most patients, the disease is manifest as ophthalmoscopically visible yellowish accumulations of protein and lipid that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane.
    Defects in C3 are a cause of susceptibility to hemolytic uremic syndrome atypical type 5 (AHUS5) [MIM:612925]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype.

  • Sequence similarities

    Contains 1 anaphylatoxin-like domain.
    Contains 1 NTR domain.

  • Post-translational
    modifications

    C3b is rapidly split in two positions by factor I and a cofactor to form iC3b (inactivated C3b) and C3f which is released. Then iC3b is slowly cleaved (possibly by factor I) to form C3c (beta chain + alpha' chain fragment 1 + alpha' chain fragment 2), C3dg and C3f. Other proteases produce other fragments such as C3d or C3g.
    Phosphorylation sites are present in the extracelllular medium.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Acylation stimulating protein cleavage product antibody
    • AHUS5 antibody
    • ARMD9 antibody
    • ASP antibody
    • C3 and PZP like alpha 2 macroglobulin domain containing protein 1 antibody
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1 antibody
    • C3 antibody
    • c3 complement antibody
    • C3adesArg antibody
    • CO3_HUMAN antibody
    • Complement C3 alpha chain antibody
    • Complement C3 antibody
    • Complement C3b alpha' chain antibody
    • Complement C3c alpha' chain fragment 1 antibody
    • Complement C3c alpha' chain fragment 2 antibody
    • Complement C3c alpha'' chain fragment 2 antibody
    • Complement C3d fragment antibody
    • Complement C3dg fragment antibody
    • Complement C3f fragment antibody
    • Complement C3g fragment antibody
    • Complement component 3 antibody
    • Complement factor 3 antibody
    • CPAMD1 antibody
    • HEL S 62p antibody
    • omplement C3 beta chain antibody
    see all

Images

  • Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    All lanes : Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 187 kDa
    Observed band size: 115,187,68 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on hepatocytes of mouse liver is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    All lanes : Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : Rat heart lysate
    Lane 8 : Rat kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 187 kDa
    Observed band size: 115,187,43,68 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2,3,5 and 6: 3 minutes; Lane 4,7 and 8: 10 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    The plasma in blood vessel of mouse cardiac muscle was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    The plasma in blood vessels of mouse cerebral cortex was positive staining.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    Western blot - Anti-C3 antibody [EPR19394] (ab200999)
    All lanes : Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution

    Lane 1 : Human serum
    Lane 2 : Human plasma
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 4 : Mouse plasma
    Lane 5 : Rat plasma
    Lane 6 : Human milk

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 187 kDa
    Observed band size: 115,187,43,68 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-5: 8 seconds; Lane 6: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on hepatocytes of rat liver is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    The plasma in blood vessels of rat kidney was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] (ab200999)

    Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    The plasma in blood vessels of rat lung was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

Publishing research using ab200999? Please let us know so that we can cite the reference in this datasheet.

ab200999 has been referenced in 13 publications.

  • Zhang LY  et al. Microglia exacerbate white matter injury via complement C3/C3aR pathway after hypoperfusion. Theranostics 10:74-90 (2020). PubMed: 31903107
  • Yuan K  et al. Complement C3 overexpression activates JAK2/STAT3 pathway and correlates with gastric cancer progression. J Exp Clin Cancer Res 39:9 (2020). PubMed: 31928530
  • Dash B  et al. Murine T Cell Maturation Entails Protection from MBL2, but Complement Proteins Do Not Drive Clearance of Cells That Fail Maturation in the Absence of NKAP. J Immunol 203:408-417 (2019). PubMed: 31175160
  • Ye J  et al. The mTORC1 signaling modulated by intracellular C3 activation in Paneth cells promotes intestinal epithelial regeneration during acute injury. Int Immunopharmacol 67:54-61 (2019). PubMed: 30530169
  • Chen SY  et al. EP4 Antagonist-Elicited Extracellular Vesicles from Mesenchymal Stem Cells Rescue Cognition/Learning Deficiencies by Restoring Brain Cellular Functions. Stem Cells Transl Med N/A:N/A (2019). PubMed: 30891948
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-C3 antibody [EPR19394]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 23°C
Fixative
Formaldehyde
Read More

Ms. Linh Hellmers

Verified customer

Submitted May 19 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-C3 antibody [EPR19394]

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA ph 9
Specification
brain tissue
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde
Read More

Herr Dr. Markus Kipp

Verified customer

Submitted Aug 21 2018

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