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    c3-antibody-epr19394-ab200999.pdf

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Immunology Innate Immunity Complement Classical Pathway
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RecombinantRabMAb

Recombinant Anti-C3 antibody [EPR19394] (ab200999)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (5) Submit a question References (29)

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

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

Conjugates logo Related conjugates and formulations

Carrier Free

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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. This information is proprietary to Abcam and/or its suppliers.

  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19394
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Complement
    • Classical Pathway
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Synapse marker
    • Signal Transduction
    • Antibodies
    • c3

Associated products

  • Alternative Versions

    • Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (ab225539)
    • Anti-C3b / iC3b antibody [7C12] (ab231078)
    • Anti-C3b / iC3b antibody [1H8] (ab231080)
    • Anti-C3 antibody [EPR19394] - BSA and Azide free (ab271967)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human C3 protein (ab157985)

Applications

The Abpromise guarantee

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 (4)
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).
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.
  • Target information above from: UniProt accession P01024 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 718 Human
    • Entrez Gene: 12266 Mouse
    • Entrez Gene: 24232 Rat
    • Omim: 120700 Human
    • SwissProt: P01024 Human
    • SwissProt: P01027 Mouse
    • SwissProt: P01026 Rat
    • Unigene: 529053 Human
    • Unigene: 19131 Mouse
    • Unigene: 11378 Rat
    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.

  • Anti-C3 antibody [EPR19394] (ab200999)
    Anti-C3 antibody [EPR19394] (ab200999)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (29)

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

ab200999 has been referenced in 29 publications.

  • Huang L  et al. L-3-n-Butylphthalide improves synaptic and dendritic spine plasticity and ameliorates neurite pathology in Alzheimer's disease mouse model and cultured hippocampal neurons. Mol Neurobiol 58:1260-1274 (2021). PubMed: 33146400
  • Li J  et al. A pentapeptide enabled AL3810 liposome-based glioma-targeted therapy with immune opsonic effect attenuated. Acta Pharm Sin B 11:283-299 (2021). PubMed: 33532193
  • Gómez Delgado I  et al. Complement Genetic Variants and FH Desialylation in S. pneumoniae-Haemolytic Uraemic Syndrome. Front Immunol 12:641656 (2021). PubMed: 33777036
  • Frey AM  et al. Insight into the human pathodegradome of the V8 protease from Staphylococcus aureus. Cell Rep 35:108930 (2021). PubMed: 33826899
  • He JH  et al. Differential and paradoxical roles of new-generation antidepressants in primary astrocytic inflammation. J Neuroinflammation 18:47 (2021). PubMed: 33602262
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-4 of 4 Abreviews or Q&A

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

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (palatine tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
palatine tonsil
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 03 2020

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

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 03 2020

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

Excellent
Abreviews
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
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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