Overview

  • Product name
    Anti-Hemopexin antibody [04]
    See all Hemopexin primary antibodies
  • Description
    Mouse monoclonal [04] to Hemopexin
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length native protein: Hemopexin purified from human plasma. The immunogen has been adsorbed onto Al(OH)3 prior to injection in mice.

  • General notes


    The fusion partner for this antibody is SP2mIL6.

Properties

Applications

Our Abpromise guarantee covers the use of ab27710 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
ELISA 1/10000.
WB Use at an assay dependent dilution.

Target

  • Function
    Binds heme and transports it to the liver for breakdown and iron recovery, after which the free hemopexin returns to the circulation.
  • Tissue specificity
    Expressed by the liver and secreted in plasma.
  • Sequence similarities
    Belongs to the hemopexin family.
    Contains 5 hemopexin-like domains.
  • Post-translational
    modifications
    N- and O-glycosylated. O-glycosylated with core 1 or possibly core 8 glycans.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Beta 1B glycoprotein antibody
    • Beta-1B-glycoprotein antibody
    • FLJ56652 antibody
    • HEMO antibody
    • HEMO_HUMAN antibody
    • Hemopexin antibody
    • Hpx antibody
    • HX antibody
    see all

References

ab27710 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question

Hi, i'm responding the questionary 1) Abcam product code ab: ab13413, ab27710, ab77811 2) Abcam order reference number or product batch number 3) Description of the problem: no bands 4) Sample preparation: Type of sample: HepG2 whole cell lysates Species : Human Lysis buffer : 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 5% glycerol Protease inhibitors: aprotinin, leupeptin, bestatin, pesptatin and E-64 Phosphatase inhibitors : Reducing agent : DTT Boiling for ≥5 min? yes Protein loaded ug/lane: generally 8 ug Positive control : Negative control : 5) Percentage of gel: 12% Type of membrane: Nitrocelulose Protein transfer verified: Panceau Red Blocking agent and concentration: I get no bands with non fat milk 5% + TBST, and some ghosty bands with BSA sigma 5% + TBST Blocking time: 2h for milk and overnight for BSA Blocking temperature: 4 oC 6) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution generally 1:1000 Diluent buffer TBST + blocking agent Incubation time O.N. utilizing milk 5% as blocking agent and 2h utilizing BSA 5% as blocking agent. Incubation temperature: 4 oC 7) Secondary antibody: anti-mouse Species: Reacts against: Concentration or dilution , generally 1:4000 Diluent buffer TBST + blocking agent Incubation time 2h Incubation temperature: 4 oC Fluorochrome or enzyme conjugate: HRP 8) Washing after primary and secondary antibodies: Buffer TBST Number of washes 3x (10 min) 9)Detection method ECL 10) How many times have you run this staining? 3 Do you obtain the same results every time? yes What steps have you altered to try and optimize the use of this antibody? Change the blocking agent (Milk to BSA).

Read More
Answer

Thank you for getting back to me with those details. Having reviewed your protocol there are a few things I may be able to suggest. The dilution for ab13413 is recommended in the range of 5-10µg/ml and ab27710 at an assay dependent dilution. I would therefore suggest decreasing the dilution that you are using trying 1/200 and 1/500 to see if any improvement is observed. Even though ab77811 is recommended at 1/1000 I would also suggest trying this with 1/200 and 1/500 as this may also need optimising with your sample. I would also suggest increasing the amount of protein that you are loading on the gel, we would normally recommend loading between 20-40 µg per well. As you are observing some bands with blocking using BSA I would continue to use this blocking agent. Initially using 3% in TBST incubating for 1-2 hours at room temperature. I would then incubate the primary antibody diluted in 1% BSA TBST for 1 hour at room temperature and overnight at 4°C with agitation. It may be that the conditions that you have been using has been over blocking the membrane. I would also reduce the wash step to 3x5 min. I realise this is alot to try with three different antibodies so I would recommend optimising each one at a time to find the best experimental conditions for each antibody. May I ask what secondary antibodies you have been using to detect the primary antibodies. ab13413, ab2771 and ab77811 are mouse IgG1, IgG2b and IgM respectively and would need to be used with secondary antibodies which are able to recognise these isotypes. I hope these suggestions help to improve the results you have been observing. If you continue to have problems please do let me know.

Read More

Answer

Thank you for contacting us. I am sorry that you are experiencing problems with these antibodies. In order to assist you as best I can could you please fill in the questionnaire I have attached to this email. This will allow me to understand more clearly the experiments that you have been performing and identify any advice I can give in order to achieve better results. It would be helpful if you did this for each antibody individually and if you could provide me with any images of the blots you have obtained this would also be of help. I look forward to hearing back from you.

Read More

Question
Answer

Thank you for your enquiry and your patience. I apologize for the delay in responding to you. I was awaiting confirmation of the information. ab27710 and ab27711 have been paired together in an ELISA. We recommend using ab27710 as the coating antibody and biotinylated ab27711 as the detection antibody. We do not sell a biotinylated version of ab27711, but there are many commercially available kits that you can use to accomplish this. We do not have any findings (it is not tested), whether ab14175 and ab14176 can be paired in a sandwich ELISA. If you require further information, please do not hesitate to contact me.

Read More

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

Sign up