Overview

  • Product name
    Anti-Factor H antibody [OX-24]
    See all Factor H primary antibodies
  • Description
    Mouse monoclonal [OX-24] to Factor H
  • Host species
    Mouse
  • Specificity
    ab118820 recognizes the Human serum complement protein factor H and a 43-49kD truncated form of factor H present at.
  • Tested applications
    Suitable for: WB, IP, ELISA, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Purified Human Factor H protein.

  • Positive control
    • Human liver tissue.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Monoclonal
  • Clone number
    OX-24
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab118820 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. Predicted molecular weight: 139 kDa.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 20 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration. PubMed: 25254972

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

  • ab118620, at 20 µg/ml, staining Factor H in formalin fixed, paraffin embedded Human liver tissue by Immunohistochemistry followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen.

References

This product has been referenced in:
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Q&A

Question

Please find my answers in the questionnaire.
Thanks


1) Abcam product code: ab 118820 Lot: GR19604-5
2) Abcam order reference number: ??????
3) Description of the problem:
The anti-GPCR GPR49 antibody showed no staining at all. Our technicians are highly experienced in establishing new antibodies (having far over 1000 antibodies established in the last years). Lot-Nr. GR19604-5
4) Sample preparation:
Species : different murine and human tissues
Type of sample: formalin fixed paraffin embedded sections
Sample preparation:
Positive control: murine gut, brain, human plazenta
Negative control Multitissue-Array with human normal and tumor samples
5) Fixation step
Yes
If yes: 10% buffered Formalin
Fixation time: several days
Fixation temperature: room temperature
6) Antigen retrieval method
HIER with EDTA-Buffer pH8, Citrate-Buffer pH6,0, different intensities
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration: Detergents in commercial LEICA-Washing-buffer
8) Blocking agent (eg BSA, serum…):
Concentration
Blocking time
Blocking temperature
9) Endogenous peroxidases blocked?
Endogenous biotins blocked?
LEICA- Refine-DAB-Kit
10) Primary antibody (If more than one was used, describe in additional notes) :
Concentration or dilution 1:25
Diluent buffer LEICA-Diluent
Incubation time 30min.
11) Secondary antibody:
Species:
LEICA-Refine DAB-Kit against Rabbit primary antibodies
12) Washing after primary and secondary antibodies:
Buffer: LEICA BondMax
Number of washes
13) Detection method
LEICA Refine-DAB-Kit on BondMax, according to the manufacturers protocols
14) How many times have you run this staining? Several times
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody?
Several Epitope Retrievals, Different Antibody-Concentrations

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Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I appreciate the experience of your team and the time they have spent on these experiments in the laboratory. I fully understand your concerns and it is disappointing the results have not been successful.

I would like to reassure you that ab118820 is tested and covered by our 6 month guarantee in IHC-P and human samples. We are happy to offer a refund, credit note or free of charge replacement when a product is not working in a successfully tested applications or species within the guarantee period. However, we do often find that suggesting some scientifically thought out optimisation tips helps to improve the results. So I hope you can understand that we like to offer the best technical support possible to provide a satisfactory outcome.

Reviewing the details, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some further details to help with my investigation of this case:

1. The secondary details state 'LEICA-Refine DAB-Kit against Rabbit primary antibodies' . The ab118820 Factor H antibody is a mouse monoclonal IgG1 primary antibody, and will require an anti mouse secondary antibody for detection. An anti-rabbit secondary will not work for detection of ab118820. Please confirm further details of the secondary antibody.

2. I can suggest to try overnight at 4oC. This often provides more efficient and specific staining.

3. Could you confirm if different times of antigen retrieval have been tried? This can sometimes require some optimization. For example, try 2, 5, 10 and 15 minutes?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested information and details of how you would like to proceed.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, IP, ELISA, IHC-P, Flow Cytometry and in human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have enclosed the IHC form as I think this is what you may be using? Please let me know if you are using a different application and I will be pleased to send you the relevant form.

In addition to this:

1. Please confirm the order number and date of purchase? Was this Abcam order ref #### purchased 14th March? Purchase order #####?

2. Regrettably, I can confirm this antibody is not tested in mouse. All tested species covered by the guarantee are listed on the online datasheets. I have reviewed the alignment of the full length human protein which has been used as an immunogen with the mouse protein, and this has only 61% alignment to mouse, which is quite low. I would therefore suggest the antibody would probably not react with mouse.

3. With regards to suggesting a different antibody to try, I would appreciate if you could confirm confirm which application you are using so I can ensure to select a suitable one for you that is covered by the guarantee.

I would appreciate if you could also provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details Antibody code:
Lot number:
Purchase order number or preferably Abcam order number:
General Information Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, low signal, non-specific staining etc.)
Sample (Species/Tissue/Cell Type/Cell Line etc.)
Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
Permeabilization step
Blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method
Positive and negative controls used (please specify)
Optimization attempts (problem solving) How many times have you tried the IHC?
Have you run a "No Primary" control? Yes No
Do you obtain the same results every time? Yes No
What steps have you altered?
Additional Notes
We would appreciate if you are also able to provide and image which would help us to assess the results

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Question

1) Abcam product code: ab 118820
2) Abcam order reference number: x

3) Description of the problem:

The Anti-Factor H antibody shows no specific staining. The positive control (liver tissue) is also negative (Pictures attached). Our technicians are highly experienced in establishing new antibodies (having far over 1000 antibodies established in the last years). The other ordered and newly established Anti-Carboxypeptidase M antibody works fine.

4) Sample preparation:
Species : TMA with liver, lymph node, lung, uterus, testis, kidney, prostate
Type of sample: formalin fixed paraffin embedded sections
Sample preparation:
Positive control: liver tissue
Negative control

5) Fixation step
Yes
If yes: 10% buffered Formalin
Fixation time: several days
Fixation temperature: room temperature

6) Antigen retrieval method
HIER with EDTA-Buffer pH8, Citrate-Buffer pH6,0, different times
PIER: Proteinase K different times

7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration: Detergents in commercial LEICA-Washing-buffer

8) Blocking agent (eg BSA, serum…):
Concentration
Blocking time
Blocking temperature

9) Endogenous peroxidases blocked?
Endogenous biotins blocked?
LEICA- Refine-DAB-Kit

10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution 1:100
Diluent buffer LEICA-Diluent
Incubation time 30min.

11) Secondary antibody:
Species:
LEICA-Refine DAB-Kit against Rabbit and Mouse primary antibodies

12) Washing after primary and secondary antibodies:
Buffer: LEICA BondMax
Number of washes

13) Detection method

LEICA Refine-DAB-Kit on BondMax, according to the manufacturers protocols

14) How many times have you run this staining? 3times
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? Different Pretreatment, Antibody-Concentrations

Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.

Read More
Answer

Vielen Dank für Ihre Antwort und für diese weiteren Informationen.

Es tut mir leid zu hören, dass der Factor H-Antikörper ab118820nicht so funktioniert hat, wie auf dem Datenblatt beschrieben (d.h. in einer Konzentration von 20 µg/ml nach HIER mit Citratpuffer pH6). Die Protokolle mit den unterschiedlichen Demaskierungsmethoden, die Sie verwendet haben,sind demProtokoll, nach demwir die Antikörper testen (s. Anhang),in der Tat sehr ähnlich. Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben und möchte Ihnen daher einen Ersatz oder eineGutschrift anbieten:

Möchten Sie noch einmal ein Röhrchen von ab118820, oder benötigen Sie einen anderen primären Antikörper?

Oder würden Sie eine Gutschrift bevorzugen?

Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Ich freue mich bald wieder von Ihnen zu hören.

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Question
Answer

Vielen Dank für Ihren Anruf.

Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.

Wie am Telefon besprochen, sende ich Ihnen hier unseren Fragebogen im Anhang. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.

Ich freue mich, bald wieder von Ihnen zu hören.

Read More

Answer

Thank you for contacting us.
Both antibodies are derived from the same clone, OX-24. Not only is one conjugated, the buffer components are slightly different as well. However, even if clones do sometimes show different behaviour regarding reactivity and application form, most of the time they react identical. And yes, ab112197 hasn't been tested in IHC yet.
If you are interested in doing so, I could make you a special offer in this case.
Please do not hesitate to contact us if you need any more advice or information.

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