Overview

  • Product name
  • Description
    Sheep polyclonal to Factor H
  • Host species
    Sheep
  • Tested applications
    Suitable for: WB, IHC-P, Double Immunodiffusion, RID, Immunoelectrophoresis, Other, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Horse, Guinea pig, Cat, Dog, Human
    Does not react with: Rabbit, Chicken, Pig
  • Immunogen

    Human 155 kDa Factor H purified from plasma.

  • General notes


    Factor H is mainly synthesised in the liver but also in macrophages and endothelium. It is primarily aplasma glycoprotein but is also found in platelets and there is a membrane bound form on some leukocytes. Consisting of a single polypeptide, the major form of Factor H has a molecular weight of 155kDa. There are two truncated forms, a non-glycosylated 49 kDa form and a glycosylated 39-43 kDaform. Plasma concentrations are in the range 200-600mg/L for the 155 kDa form and 1-5mg/L for thetruncated forms. Factor H is a major regulatory protein of the complement system. By binding to C3b it either displacesor prevents the binding of Bb (activated Factor B). When bound to Factor H, C3b is susceptible tocleavage by Factor 1 to yield iC3b. Factor H is released or modified following this cleavage. The regulatory role of Factor H is essential because C3bBb is not only a C5 convertase but a C3 convertaseand so has a positive feedback effect, potentially consuming the entire C3 pool if unregulated.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituents: 0.1% EACA, 0.01% Benzamidine, 0.0292% EDTA
  • Concentration information loading...
  • Purity
    IgG fraction
  • Purification notes
    Antiserum is prepared by immunisation of sheep with human Factor H and, if necessary, adsorption tomonospecificity by use of solid-phase adsorbents. An immunoglobulin fraction is then produced. The titre is adjusted so that inter-batch variation is within 10%. The product is finally 0.2µm filtered.
  • Primary antibody notes
    Factor H is mainly synthesised in the liver but also in macrophages and endothelium. It is primarily aplasma glycoprotein but is also found in platelets and there is a membrane bound form on some leukocytes. Consisting of a single polypeptide, the major form of Factor H has a molecular weight of 155kDa. There are two truncated forms, a non-glycosylated 49 kDa form and a glycosylated 39-43 kDaform. Plasma concentrations are in the range 200-600mg/L for the 155 kDa form and 1-5mg/L for thetruncated forms. Factor H is a major regulatory protein of the complement system. By binding to C3b it either displacesor prevents the binding of Bb (activated Factor B). When bound to Factor H, C3b is susceptible tocleavage by Factor 1 to yield iC3b. Factor H is released or modified following this cleavage. The regulatory role of Factor H is essential because C3bBb is not only a C5 convertase but a C3 convertaseand so has a positive feedback effect, potentially consuming the entire C3 pool if unregulated.
  • Clonality
    Polyclonal
  • Myeloma
    unknown
  • Isotype
    IgG
  • Light chain type
    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8842 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
WB Use at an assay dependent concentration. PubMed: 20813971
Suitability for use in nephelometry and enzyme-linked immunosorbent assays have not been assessed but use in such assays should not necessarily be excluded.
IHC-P Use at an assay dependent concentration. PubMed: 20813971
Double Immunodiffusion Use at an assay dependent concentration. 10µL antiserum vs 10µL plasma
RID Use at an assay dependent concentration. (and Rocket IEP) 1µL antiserum/cm² gel vs 5µL neat - 1/10 dilution human plasma.
The use of 3% PEG 6000 with 1.2% agarose in a suitable buffer (such as TBE or Tris-barbital pH >8.2) is recommended.
Immunoelectrophoresis Use at an assay dependent concentration. 100µL antiserum vs 5µL plasma
Other Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration. PubMed: 22461909

Target

  • Function
    Factor H functions as a cofactor in the inactivation of C3b by factor I and also increases the rate of dissociation of the C3bBb complex (C3 convertase) and the (C3b)NBB complex (C5 convertase) in the alternative complement pathway.
  • Tissue specificity
    Expressed by the liver and secreted in plasma.
  • Involvement in disease
    Genetic variations in CFH are associated with basal laminar drusen (BLD) [MIM:126700]; also known as drusen of Bruch membrane or cuticular drusen or grouped early adult-onset drusen. Drusen are extracellular deposits that accumulate below the retinal pigment epithelium on Bruch membrane. Basal laminar drusen refers to an early adult-onset drusen phenotype that shows a pattern of uniform small, slightly raised yellow subretinal nodules randomly scattered in the macula. In later stages, these drusen often become more numerous, with clustered groups of drusen scattered throughout the retina. In time these small basal laminar drusen may expand and ultimately lead to a serous pigment epithelial detachment of the macula that may result in vision loss.
    Defects in CFH are the cause of complement factor H deficiency (CFH deficiency) [MIM:609814]. CFH deficiency determines uncontrolled activation of the alternative complement pathway with consumption of C3 and often other terminal complement components. It is associated with a number of renal diseases with variable clinical presentation and progression, including membranoproliferative glomerulonephritis and atypical hemolytic uremic syndrome. CFH deficiency patients may show increased susceptibility to meningococcal infections.
    Defects in CFH are a cause of susceptibility to hemolytic uremic syndrome atypical type 1 (AHUS1) [MIM:235400]. 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.
    Genetic variation in CFH is associated with age-related macular degeneration type 4 (ARMD4) [MIM:610698]. 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 (known as drusen) that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane.
  • Sequence similarities
    Contains 20 Sushi (CCP/SCR) domains.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • adrenomedullin binding protein antibody
    • age related maculopathy susceptibility 1 antibody
    • AHUS 1 antibody
    • AHUS1 antibody
    • AMBP 1 antibody
    • AMBP1 antibody
    • ARMD 4 antibody
    • ARMD4 antibody
    • ARMS 1 antibody
    • ARMS1 antibody
    • beta 1 H globulin antibody
    • beta 1H antibody
    • beta1H antibody
    • CFAH_HUMAN antibody
    • CFH antibody
    • CFHL 3 antibody
    • CFHL3 antibody
    • Complement factor H antibody
    • complement factor H, isoform b antibody
    • Factor H antibody
    • factor H like 1 antibody
    • FH antibody
    • FHL 1 antibody
    • FHL1 antibody
    • H factor 1 (complement) antibody
    • H factor 1 antibody
    • H factor 2 (complement) antibody
    • HF 1 antibody
    • HF 2 antibody
    • HF antibody
    • HF1 antibody
    • HF2 antibody
    • HUS antibody
    • MGC88246 antibody
    see all

Images

  • All lanes : Anti-Factor H antibody (ab8842) at 1/500 dilution

    Lane 1 : Purified Human Factor H
    Lane 2 : Human serum sample (albumin depleted)
    Lane 3 : Mouse serum sample (albumin depleted)
    Lane 4 : Rat serum sample (albumin depleted)
    Lane 5 : Guinea pig serum sample (albumin depleted)
    Lane 6 : Cattle serum sample (albumin depleted)
    Lane 7 : Horse serum sample (albumin depleted)
    Lane 8 : Dog serum sample (albumin depleted)
    Lane 9 : Cat serum sample (albumin depleted)

    Developed using the ECL technique.

    Performed under non-reducing conditions.


    Samples were fractionated by non-reducing SDS-PAGE (10 µg/well/animal species) and transferred to nitrocellulose membranes. Membranes were blocked overnight at 4°C in SuperBlock buffer and then incubated for 2 hours (37°C with shaking) with ab8842 diluted in TTBS buffer [10 mM Tris/HCl (pH 8.3), 0.05% Tween-20 and 150 mM NaCl] with 1% skim milk. Membranes were washed 3 times with TTBS, incubated with rabbit anti-sheep HRPO antibody diluted to 1/400,000 in TTBS with 1% skim milk for 1 hour (37°C with shaking) and then washed 3 times.

References

This product has been referenced in:
  • Li TR  et al. The role of the microRNA-146a/complement factor H/interleukin-1ß-mediated inflammatory loop circuit in the perpetuate inflammation of chronic temporal lobe epilepsy. Dis Model Mech 11:N/A (2018). Read more (PubMed: 29590637) »
  • Marazita MC  et al. Oxidative stress-induced premature senescence dysregulates VEGF and CFH expression in retinal pigment epithelial cells: Implications for Age-related Macular Degeneration. Redox Biol 7:78-87 (2016). Read more (PubMed: 26654980) »
See all 13 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thanks so much for all your troubleshooting efforts and for including the protocol details and IHC images in the attachement!
I agree this antibody is not working as expected. I would be happy to offer a replacement, credit or refund.
In your reply, please include the Abcam order reference number or your internal PO number associated with the purchase of ab8842 so I can process your request.

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Answer

DISCOUNT CODE:xx
Expiration date: xx

I am very pleased to hear you would like to accept our offer and test ab8842 in cow. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forcow and include this code in the Additional Comments section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

I will send you the invoice proforma shortly as well.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for submitting your completed questionnaire. I'm sorry to hear this vial of ab8842 is giving you trouble.
To decrease background bands, you might try further dilution of the secondary, to 1:10000, for example. In addition, it might be helpful to look at the following reference, which has used this antibody in western blotting on a number of tissue lysates and is found under the 'Specific references' tab on our online product datasheet:
Bhide MR et al. Complement factor H binding by different Lyme disease and relapsing fever Borrelia in animals and human. BMC Res Notes 2:134 (2009). WB; Human, Mouse, Rat, Cat, Cow, Dog, Guinea pig, Horse. PubMed: 19604355
For example, they note using non-reducing conditions for their blots.
I hope this is helpful. I look forward to hearing if your results improve.

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Answer

Thank you very much for your inquiry and patience while I checked this information.

Indeed, the laboratory has tested this antibody ab8842 via Ouchterlony (double immunodiffusion). In this assay they have not detected any crossreactivity with Cow.

Western blot is however a more sensitive detection method, and it seems that this antibody is working well in Western blot also on bovine samples.The image is from Bhide MR et al, BMC Res Notes. 2009 Jul 15;2:134, Fig 1.

I would therefore think that the antibody will work for your purpose. In order however to provide you with some sort of guarantee, I would propose to offer you a testing discount. This would provide you with a second antibody for free after feedback of your experience with this antibody via an Abreview. It would also povide us with a secondand more specific feedback on the use of this antibody to detect the Cow protein.

The details of the testing discount I can offer you are here below:

I can offer a discount off a future purchase if you buy ab8842 now, test it incow and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab8842 in cow. I will then send a discount code. This code must be issued before purchasing ab8842 so please wait for my reply before ordering.

2. Purchase ab8842 either by phone, fax, or online (www.abcam.com).

3. Test it in cow.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab8842 turns out to be unsuitable for cow, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting Abcam regarding this. This antibody is produced using purified human H factor (from plasma) as the immunogen. The product is polyclonal as well. Polyclonal antibodies created by introducing the immunogen to the host species and then harvesting the antibodies that are created. As such, polyclonals are contain the antibodies to the immunogen that have been created by a number of different cells. They will also recognize a number of different epitopes. This makes them very powerful tools for recognizing proteins in different forms but as such a specific epitope cannot be mapped as it can with a monoclonal antibody. Since the polyclonal antibody was also created from the whole protein, there may be binding from numerous locations within the entire H factor sequence. Please let me know if you have questions or there are other ways that I may be able to help you.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (serum)
Specification
serum
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 17 2006

Answer

Unfortunately this antibody has not been tested with Factor H-like protein-1 (FHL-1) or any of the Factor H related proteins (FHR), we only immunise against total factor H. My feeling is that the product probably will bind FHL-1 and FHR however I cannot guarantee nor prove that this is the case. Please contact us again if you have any additional questions.

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Answer

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in IHC. All tested applications are specified on Abcam product datasheets. We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will offer you 50 Abpoints which can be redeemed on a number of rewards (a further 100 Abpoints will be offered for an image). If you have any additional questions, please contact us again.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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