Overview

  • Product name
    Anti-ABCA12 antibody
  • Description
    Rabbit polyclonal to ABCA12
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Guinea pig, Cow, Cat, Dog
  • Immunogen

    Synthetic peptide corresponding to a region within internal sequence amino acids 1979-2028 (TTIFKMLTGD IIPSSGNILI RNKTGSLGHV DSHSSLVGYC PQEDALDDLV) of Human ABCA12 (NP_056472; UniProt ID: Q86UK0 isoform 2).

  • Positive control
    • Human fetal stomach lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab98976 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 a concentration of 1 µg/ml. Predicted molecular weight: 257 kDa.Can be blocked with ABCA12 peptide (ab127886). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function
    Probable transporter involved in lipid homeostasis.
  • Tissue specificity
    Mainly expressed in the stomach, placenta, testis and fetal brain.
  • Involvement in disease
    Defects in ABCA12 are the cause of ichthyosis harlequin (HI) [MIM:242500]; also known as harlequin fetus. HI is a very severe skin disorder in which the neonate is born with a thick covering of armor-like scales. The skin dries out to form hard diamond-shaped plaques separated by fissures, resembling 'armor plating'. The normal facial features are severely affected, with distortion of the lips (eclabion), eyelids (ectropion), ears, and nostrils. Affected babies are often born prematurely and rarely survive the perinatal period.
    Defects in ABCA12 are the cause of ichthyosis lamellar type 2 (LI2) [MIM:601277]; also known as ichthyosis congenita IIB (ICR2B). LI is a non-bullous ichthyosis, a skin disorder characterized by abnormal cornification of the epidermis. It is one the most severe forms of ichthyoses apparent at birth and persisting throughout life. LI patients are born encased in a tight, shiny, translucent covering called collodion membrane. Over the first weeks of life, the collodion membrane is gradually replaced by generalized large, dark brown, plate-like scales with minimal to no erythroderma. Tautness of facial skin commonly results in ectropion, eclabium and scarring alopecia of the scalp. Common complications are severe heat intolerance and recurrent ear infections.
  • Sequence similarities
    Belongs to the ABC transporter superfamily. ABCA family.
    Contains 2 ABC transporter domains.
  • Domain
    Multifunctional polypeptide with two homologous halves, each containing an hydrophobic membrane-anchoring domain and an ATP binding cassette (ABC) domain.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ABC transporter A family member 12 antibody
    • ABC transporter ABCA.12 antibody
    • ABC12 antibody
    • ABCA12 antibody
    • ABCAC_HUMAN antibody
    • AtABCA12 antibody
    • ATH16 antibody
    • ATP binding cassette 12 antibody
    • ATP binding cassette sub family A (ABC1) member 12 antibody
    • ATP binding cassette sub family A member 12 antibody
    • ATP binding cassette transporter 12 antibody
    • ATP-binding cassette 12 antibody
    • ATP-binding cassette sub-family A member 12 antibody
    • ATP-binding cassette transporter 12 antibody
    • Ichthyosis congenita II lamellar ichthyosis B antibody
    • ICR2B antibody
    • LI2 antibody
    • Putative ABC2 homolog 16 antibody
    see all

Images

  • Anti-ABCA12 antibody (ab98976) at 1 µg/ml + Human fetal stomach lysate at 10 µg

    Predicted band size: 257 kDa



    Gel concentration: 6-18%

References

ab98976 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human primary fibroblast)
Loading amount
30 µg
Specification
human primary fibroblast
Gel Running Conditions
Reduced Denaturing (7% acrylamide gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Jian Yuan-Ren

Verified customer

Submitted Apr 12 2012

Answer

Thank you very much for your reply.

I understand the customer's frustration, however I can not offer a free of charge replacement in this case as the antibody was working as stated on the datasheet.

If the customercould submit an Abreview with his results and explaining in the "additional notes" the details of theoptimisation steps performed, we willprovide exceptionnalya free new unit of ab98976. Indeed, we can make this exception as I believe that the Abreview willavoid that other users will have the same problem.I hope this will satisfy the customer. Please do inform me as soon as possible when the Abreview has been submitted. Thank you very much.

Please let me know if you have any questions.

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Answer

Thank you for this update!


I am very happy to hear that the suggestions seemed to have resolved the problem. Unfortunately, there was no image attached to the email. Can you please send it again?

Thank you very much!

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Question

LOT NUMBER gr65334-1 ORDER NUMBER 1007542 DESCRIPTION OF THE PROBLEM Multiple bands, two bands were there SAMPLE - Species: human - What’s cell line or tissue: primary fibroblast cell (from the newborns) - Cell extract or Nuclear extract: cell extract - Purified protein or Recombinant protein: Whole cell extraction PRIMARY ANTIBODY - Species: rabbit polyclonal - Reacts against: human - At what dilution(s) have you tested this antibody: 1:1000 - What dilution buffer was used: 5% non-fat-milk in TBST - Incubation time: 1hr - Incubation temperature: room temperature - What washing steps were done: 15min x 1 + 5min x 3 DETECTION METHOD ECl+ ANTIBODY STORAGE CONDITIONS -20 SAMPLE PREPARATION - What lysis buffer was used: 20mM potassium phosphate buffer + sonication - What protease inhibitors were used: cocktail protease inhibitor - What loading buffer was used: NuPAGE LDS sample buffer (invitrogen:Cat no.NP0007) - Phosphatase inhibitors - Did you heat the samples: temperature and time: 70℃ for 10min AMOUNT OF PROTEIN LOADED 30ug ELECTROPHORESIS/GEL CONDITIONS - Reducing or non reducing gel: Reducing - Reducing agent: 2-ME - Gel percentage : 4-12% TRANSFER AND BLOCKING CONDITIONS ransfer conditions: (Type of membrane, Protein transfer verified): PVDF, Coomassie blue (Invitrogen iBlot dry blotting system Cat no. IB10001) Blocking conditions - Buffer: 5% non-fat-milk in TBST - Blocking agent: milk, BSA, serum, what percentage: 5% - Incubation time: 1hr - Incubation temperature: room temperature SECONDARY ANTIBODY - Species: goat - Reacts against: rabbit - At what dilution(s) have you tested this antibody: 1:5000 - Incubation time: 1hr - Wash steps: 15min x 1 + 5min x 3 - Fluorochrome or enzyme conjugate: enzyme conjugate - Do you know whether the problems you are experiencing come from the secondary? HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No

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Answer

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

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.

Reviewing this case, I would like to offer somecomments on the results from ab98976.
I believe that the double band (very tight double-band at the correct size) is a biological phenomenon and might indicate very interesting results for your research. In sample 49, it appears to only show a single band,whereas the other samples have double bands with different strengths.
There are multiple potential reasons for this double band:
1.) It is known that multiple isoform from this protein exist. I did not find any publication that indicated if this was ever tested in human primary fibroblasts and which one of the isoforms is expressed there. Maybe even two isoforms are expressed. The immunogen of this antibodyis present in both isoforms.
2.)ABCA12 is also a heavy glycosylated protein and the double band could suggest different glycosylation states of the protein in these samples. This can be tested by de-glycosylating the proteins before running the sample on the gel which then should only produce one band if the double band is due to glycosylation.
3.)There are also three putative phosphorylation sites in ABCA12. The phosphorylation status will also affect the bandsize of ABCA12. (Raijmakers R, et al. (2010) Exploring the human leukocyte phosphoproteome using a microfluidic reversed-phase-TiO2-reversed-phase high-performance liquid chromatography phosphochip coupled to a quadrupole time-of-flight mass spectrometer. Anal Chem 82, 824-32)

We are happy to offer this technical support. In the event that a product is not functioning in the applications 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 hope this information is helpful. Please feel free to contact me again with any further questions.

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Answer

Thank you for your enquiry. I can confirm that as the datasheet indicates ab98976 has only been tested and characterized for Western blot application. Currently, we do not have any data regarding IHC, though it does not necessary mean that ab98976 is unsuitable for this application. We would welcome any feedback on this antibody, so please consider submitting an Abreview with your results. I hope this helps and if I can assist further, please do not hesitate to contact me.  

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Answer

Thank you very much for your kind reply and the image.

Indeed, the results look very good to me. If I understood correctly, the 3rd band is where the customer used the sample with the mutated ABCA12.

The second band all on top could be the isoform 2, as described on SwissProt as well http://www.uniprot.org/uniprot/Q86UK0

The bands all at around 80kD can be either background,new isoformsor degradation products. I would recommend to use always proteinase inhibitors when preparing the samples.

If the customer would like to improve the shape of the bands, I can recommend to optimise the loading so that the protein will enter equally the gel. Points to optimise are the loading buffer and the volume loaded on the gel. If the samples in the well are not homogeneously distributed, but are towards the walls of the well, the band in the Western blot might not be a straight band.

I hope these comments are useful for the customer. Please do let me know if the customer has actually a concern with this data/antibody or if the customer has other questions.

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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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