RecombinantRabMAb

Recombinant Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

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Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Immunoprecipitation - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
  • Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23764-1] to Complement factor 8 beta/C8B - BSA and Azide free
  • Suitable for: IHC-P, WB, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free
    See all Complement factor 8 beta/C8B primary antibodies

  • Description

    Rabbit monoclonal [EPR23764-1] to Complement factor 8 beta/C8B - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
    Unsuitable for: Flow Cyt or ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: Human kidney, liver and large intestine tissue lysates; Human, mouse and rat serum; LIF-tagged human C8B recombinant protein. IHC-P: Human thyroid and liver tissue. IP: Human kidney tissue lysate.

  • General notes

    ab278049 is the carrier-free version of ab278045.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab278049 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
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).
IP
Use at an assay dependent concentration.
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).
IP
Use at an assay dependent concentration.
Application notes
Is unsuitable for Flow Cyt or ICC.

Target

  • Function

    Constituent of the membrane attack complex (MAC) that plays a key role in the innate and adaptive immune response by forming pores in the plasma membrane of target cells.

  • Involvement in disease

    Defects in C8B are a cause of complement component 8 deficiency type 2 (C8D2) [MIM:120960]. A rare defect of the complement classical pathway associated with susceptibility to severe recurrent infections, predominantly by Neisseria gonorrhoeae or Neisseria meningitidis.

  • Sequence similarities

    Belongs to the complement C6/C7/C8/C9 family.
    Contains 1 EGF-like domain.
    Contains 1 LDL-receptor class A domain.
    Contains 1 MACPF domain.
    Contains 2 TSP type-1 domains.

  • Post-translational
    modifications

    N-glycosylated; contains one or two bound glycans. Not O-glycosylated.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • C8b antibody
    • CO8B_HUMAN antibody
    • Complement component 8 beta polypeptide antibody
    • Complement component 8 subunit beta antibody
    • Complement component C8 beta chain antibody
    see all

Images

  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    All lanes : Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

    Lane 1 : Human kidney tissue lysate
    Lane 2 : Human liver tissue lysate
    Lane 3 : Human large intestine tissue lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 67 kDa
    Observed band size: 67 kDa



    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (PMID:21750743).

    Negative: human brain (PMID:16483657, PMID:27005612).

    Exposure time: 3 minutes.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human thyroid tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 µg/ml) dilution followed by a ready to use secondary antibody. Positive staining on the blood plasma in human thyroid (PMID: 9476128). The section was incubated with ab278045 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use secondary antibody.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunoprecipitation - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Immunoprecipitation - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Complement factor 8 beta/C8B was immunoprecipitated from 0.35 mg human kidney tissue lysate with ab278045 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab278045 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Human kidney tissue lysate 10 µg

    Lane 2: ab278045 IP in human kidney tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab278045 in human kidney tissue lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution + Human serum at 20 µl

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 67 kDa
    Observed band size: 67 kDa



    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (PMID:21750743).

    Exposure time: 15 seconds.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 µg/ml) dilution  followed by a ready to use secondary antibody. Cytoplasmic staining on human liver. The section was incubated with ab278045 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use secondary antibody).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    All lanes : Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

    Lane 1 : Mouse serum
    Lane 2 : Rat serum

    Lysates/proteins at 20 µl per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 67 kDa
    Observed band size: 67 kDa



    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (PMID:21750743).

    This blot was developed using a higher sensitivity ECL substrate.

    Exposure time: 26 seconds.

  • Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Western blot - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    All lanes : Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] (ab278045) at 1/1000 dilution

    Lane 1 : LIF-tagged human C8B recombinant protein, 30 ng
    Lane 2 : E.coli extracts containi his-tagged human C8A recombinant protein, 30 ng
    Lane 3 : E.coli extracts containi his-tagged human C9 recombinant protein, 30 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 67 kDa



    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This antibody has no cross reaction with human C8A and C9.

    Sample loaded onto lane 1 was purified protein extracted from HEK293 expressing C8B, which contains a LIF-tag (40 kDa). 

    C8A and C9 recombinant proteins were expressed in E.coil expression systems.

    Exposure time: 26 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

    This data was developed using ab278045, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Complement factor 8 beta/C8B with ab278045 at 1/1000 (0.533 µg/ml) dilution followed by a ready to use secondary antibody. The section was incubated with ab278045 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Negative control: no staining on human cerebrum.

    Secondary antibody only control: Secondary antibody is a ready to use secondary antibody.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)
    Anti-Complement factor 8 beta/C8B antibody [EPR23764-1] - BSA and Azide free (ab278049)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab278049? Please let us know so that we can cite the reference in this datasheet.

ab278049 has not yet been referenced specifically in any publications.

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