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AB250455

Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free

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Rabbit Recombinant Monoclonal C4B-d / C4B-b antibody. Carrier free. Suitable for ICC/IF, WB, IHC-P and reacts with Human samples.

View Alternative Names

CO4, CPAMD3, C4B_2, C4B, Complement C4-B, Basic complement C4, C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3

5 Images
Immunocytochemistry/ Immunofluorescence - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling c4b with purified ab181241 at 1/100 dilution (10 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181241).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling C4b with purified ab181241 at 1/75 dilution (9.53 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181241).

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)
  • WB

Unknown

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)

The molecular weights observed are consistent with what have been described in the literature (PMID : 15632414, 31178241 and 30210498).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181241).

All lanes:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (<a href='/en-us/products/primary-antibodies/c4b-d-c4b-b-antibody-epr11203-27-ab181241'>ab181241</a>) at 1/10000 dilution

Lane 1:

Human liver lysate at 20 µg

Lane 2:

Human heart lysate at 20 µg

Lane 3:

Human testis lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 193 kDa

Observed band size: 200 kDa,70 kDa,93 kDa

false

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)
  • WB

Unknown

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)

The molecular weights observed are consistent with what have been described in the literature (PMID : 15632414, 31178241 and 30210498).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181241).

All lanes:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (<a href='/en-us/products/primary-antibodies/c4b-d-c4b-b-antibody-epr11203-27-ab181241'>ab181241</a>) at 1/10000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Human plasma lysate at 15 µg

Lane 3:

Human serum lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 193 kDa

Observed band size: 200 kDa,70 kDa,93 kDa

false

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)
  • WB

Lab

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] - BSA and Azide free (AB250455)

This data was developed using ab181241, the same antibody clone in a different buffer formulation.

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

Lanes 1 - 5:

Western blot - Anti-C4B-d / C4B-b antibody [EPR11203-27] (<a href='/en-us/products/primary-antibodies/c4b-d-c4b-b-antibody-epr11203-27-ab181241'>ab181241</a>) at 1/1000 dilution

Lanes 6 - 10:

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/en-us/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>) at 1/5000 dilution

Lanes 1 and 6:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lanes 2 and 7:

293T cells transfected with a human C4d-A (Uniprot: P0C0L4 aa957-1336) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 3 and 8:

293T cells transfected with a human C4d-B (Uniprot: P0C0L5 aa957-1336) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 4 and 9:

293T cells transfected with a human C4-A alpha chain (Uniprot: P0C0L4 aa680- 1446) expression vector containing a His-tag whole cell lysate at 20 µg

Lanes 5 and 10:

293T cells transfected with a human C4b-A (Uniprot: P0C0L4 aa757-1446) expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 45-100 kDa

false

Exposure time: 1s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR11203-27

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IHC-P, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

This antibody was shown to detect C4d-A, C4d-B, C4b-A and the C4A alpha chain in Western Blot experiments.

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Primary Antibodies

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Primary Antibodies

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBL1+MBL2 antibody [EPR18381-156] (AB190834)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." } } }
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Product details

ab250455 is the carrier-free version of ab181241.

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The 'C4b' protein also known as the 'C4b complement' plays a significant role in the immune system. This protein is a part of the larger complement system specifically as a segment derived from the cleavage of the native 'C4 complement'. It has an estimated molecular mass of about 200 kDa. The 'C4b' protein is prominently expressed on the surface of cells and in body fluids where the immune response is active. As an important player in the complement pathway it binds covalently to foreign particles labeling them for destruction or removal by immune cells.
Biological function summary

The 'C4b complement' interacts with various other components of the immune system. It is particularly involved in the formation of the C3 and C5 convertase complexes which are essential for the opsonization and activation cascade leading to pathogen elimination. Its attachment to cellular surfaces provides a docking point for proteins such as C2 facilitating the further cleavage and progression of the complement cascade. This interaction is critical for driving the classical and lectin pathways of complement activation.

Pathways

The 'C4b protein' is integral to the classical pathway of complement activation as well as to the lectin pathway. It associates with proteins like C1s C1q and factor B forming complexes that execute various stages of immune defenses. By forming C3 and C5 convertase 'C4b' not only promotes phagocytosis but also inflammation and cell lysis through membrane attack complexes. These processes underlie the body's ability to address microbial invasions efficiently.

'C4b complement dysregulation' can lead to autoimmune conditions such as systemic lupus erythematosus (SLE) and type 1 diabetes. In SLE reduced or malfunctional 'C4b' increases susceptibility to infections and triggers inappropriate immune responses. It can also correlate with the deficiency of related proteins like C2 which further impairs the complement system. These associations indicate that proper regulation of 'C4b' is essential for maintaining immune system stability and preventing overactive immune responses.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Precursor of non-enzymatic components of the classical, lectin and GZMK complement pathways, which consist in a cascade of proteins that leads to phagocytosis and breakdown of pathogens and signaling that strengthens the adaptive immune system.. Complement C4b-B. Non-enzymatic component of C3 and C5 convertases (By similarity). Generated following cleavage by complement proteases (C1S, MASP2 or GZMK, depending on the complement pathway), it covalently attaches to the surface of pathogens, where it acts as an opsonin that marks the surface of antigens for removal (By similarity). It then recruits the serine protease complement C2b to form the C3 and C5 convertases, which cleave and activate C3 and C5, respectively, the next components of the complement pathways (PubMed : 8538770). Complement C4b-B isotype catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens, while C4b-A isotype is responsible for effective binding to form amide bonds with immune aggregates or protein antigens (PubMed : 8538770).. C4a anaphylatoxin. Putative humoral mediator released following cleavage by complement proteases (C1S, MASP2 or GZMK, depending on the complement pathway). While it is strongly similar to anaphylatoxins, its role is unclear. Was reported to act as a mediator of local inflammatory process; however these effects were probably due to contamination with C3a and/C5a anaphylatoxins in biological assays.
See full target information C4B
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Product promise

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For full details, please see our Terms & Conditions

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