Overview

  • Product name
    Anti-Apolipoprotein B antibody
    See all Apolipoprotein B primary antibodies
  • Description
    Rabbit polyclonal to Apolipoprotein B
  • Host species
    Rabbit
  • Specificity
    Cross-reacts with Apo B48 and B100.
  • Tested applications
    Suitable for: IHC-Fr, IP, ELISA, IHC-P, Immunodiffusion, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length native protein (purified) corresponding to Mouse Apolipoprotein B. Apolipoprotein B48 purified from pooled mouse plasma very low-density lipoprotein.

Applications

Our Abpromise guarantee covers the use of ab20737 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
IHC-Fr 1/1000.
IP Use at an assay dependent concentration. PubMed: 18497885
ELISA Use at an assay dependent concentration. PubMed: 21668821
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunodiffusion Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 549 kDa.

Target

  • Function
    Apolipoprotein B is a major protein constituent of chylomicrons (apo B-48), LDL (apo B-100) and VLDL (apo B-100). Apo B-100 functions as a recognition signal for the cellular binding and internalization of LDL particles by the apoB/E receptor.
  • Involvement in disease
    Hypobetalipoproteinemia, familial, 1
    Familial ligand-defective apolipoprotein B-100
    Defects in APOB associated with defects in other genes (polygenic) can contribute to hypocholesterolemia.
  • Sequence similarities
    Contains 1 vitellogenin domain.
  • Post-translational
    modifications
    Palmitoylated; structural requirement for proper assembly of the hydrophobic core of the lipoprotein particle.
  • Cellular localization
    Cytoplasm. Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apo B-100 antibody
    • Apo B-48 antibody
    • ApoB 100 antibody
    • ApoB 48 antibody
    • APOB antibody
    • APOB_HUMAN antibody
    • Apolipoprotein B (including Ag(x) antigen) antibody
    • Apolipoprotein B 100 antibody
    • Apolipoprotein B antibody
    • Apolipoprotein B-48 antibody
    • Apolipoprotein B48 antibody
    • FLDB antibody
    • LDLCQ4 antibody
    see all

Images

  • ab20737 staining Apoplipoprotein B in murine intestinal tissue by Immunohistochemistry (frozen sections). Tissue was fixed with 10% formalin and permeabilized using Triton X-100. Samples were then blocked with 10% serum for 30 minutes at 24°C followed by incubation with the primary antibody, at a 1/100 dilution, for 16 hours at 4°C. A donkey anti-rabbit F(ab')2 fragment conjugated to Cy5® was used as secondary antibody at a 1/1000 dilution.

    See Abreview

  • ab20737 at a 1/100 dilution staining Apolipoprotein B in murine small intestinal tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed. Samples were then permeabilized using 0.2% PBST and blocked using 20% serum. An Alexa Fluor® 488 conjugated Goat anti-rabbit IgG was used as the secondary at a 1/100 dilution.
  • ab20737 staining Apolipoprotein in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% neutral buffered formalin and blocked with 1.5% serum for 45 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (8.5ug/ml in 0.3% Triton X-100 in PBS) for 1 hour at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (7ug/ml) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Dong Z  et al. Loss of LCAT activity in the golden Syrian hamster elicits pro-atherogenic dyslipidemia and enhanced atherosclerosis. Metabolism 83:245-255 (2018). WB . Read more (PubMed: 29526535) »
  • Sun H  et al. PCSK9 deficiency reduces atherosclerosis, apolipoprotein B secretion, and endothelial dysfunction. J Lipid Res 59:207-223 (2018). Read more (PubMed: 29180444) »
See all 34 Publications for this product

Customer reviews and Q&As

1-10 of 26 Abreviews or Q&A

Answer


Below is information regarding the use of ab20737 in western blot.

1. Working dilution for immunoblotting - start low 1:100 - 1:500.
2. Molecular Weight of Apo B100 is 500kDa; Apo B48 is 250kDa. The western blot was run against purified LDL from mouse and against human plasma. Also tested mouse VLDL to detect ApoB48. When testing whole plasma, you may see additional bands (possible proteolytic fragments of ApoB100 and B48).
3. Use 1% BSA for blocking in the immunoblot part of the procedure.

Our general recommendations for transfer of large proteins, and a recommended paper that outlines a protocol for large protein transfer is given in our western blot guide below. As I mentioned, I recommended ensuring there is no methanol in your transfer buffer and including 0.1% SDS in the transfer buffer as well.

https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf

Below is a link to a paper that used ab20737 in western blot with mouse samples.

http://www.ncbi.nlm.nih.gov/pubmed/17438339?dopt=Abstract

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Whole liver)
Gel Running Conditions
Reduced Denaturing (5% gel, run at 150V for 1.5 hours.)
Loading amount
40 µg
Specification
Whole liver
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 20 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Specification
Kidney
Fixative
10% neutral buffered formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0 (20 min, 93*C)
Permeabilization
No
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jan 24 2013

Answer

Because we carry over 100,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If a species or application is untested you can use our products without financial risk by participating in our AbTrial program.

You would have to buy the ab117317 but once you submitted the Abreview you will receive a credit over the whole value of ab117317. You will receive this for either a positive and negative result of the testing.

I hope this information is helpful. Please let me know if you like to except the AbTrial offer and I would be happy to issue the Trial Code for you. If you have further questions please do not hesitate to contact me.

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Answer

Thank you for your inquiry.

I am happy to confirm that we offer an Anti-Apolipoprotein B antibody that is tested and guaranteed for ICC and WB in rat, human and mouse samples:

ab117317b (Anti-Apolipoprotein B antibody)

https://www.abcam.com/index.html?datasheet=117317 (or use the following: https://www.abcam.com/index.html?datasheet=117317).

ab117317 is a chicken IgY antibody a suitbale secondary antibody is anti chicken IgY.

For example:

ab97135

https://www.abcam.com/index.html?datasheet=97135 (or use the following: https://www.abcam.com/index.html?datasheet=97135).

Apolipoprotein B is a very big protein (over 500kDa plus many modifications) and I would suggest to review these tips for big proteins in WB.

The following reference discusses a gel and buffer system that allows transfer of proteins as large as 500 kD:

Bolt and Mahoney, High-efficiency blotting of proteins of diverse sizes following sodium dodecyl sulfate–polyacrylamide, gel electrophoresis. Analytical Biochemistry 247, 185–192 (1997).

https://www.abcam.com/index.html?pageconfig=resource&rid=11404#C2

I hope this information is helpful and wish you good luck with your experiments.

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Answer

Thank you for contacting us. Your credit note ID is xxxxx. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

Thank you for your reply.

I can gladly create this credit note/refund for you provided that you have purchase this item within the last six months. In order for me to issue this could you please provide me the Abcam order confirmation number for purchase of ab20737.

I look forward to your reply. Please let me know if you have any other questions.

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Question
Answer

Thank you for your reply. If your Abcam order was placed by credit card, the credit note indicates the dollar amount that will be refunded to your credit card account. Please allow 2-5 business days for the credit to process and appear on your statement. If your Abcam order was placed by Purchase Order andthe original invoice has not yet been paid to Abcam, your credit note will be forwarded on to your institution’s accounts payable department. They will subtract the credit amount from the invoice total, and disburse a check (if any) for the correct amount payable to Abcam. If your Abcam order was placed by Purchase Order andthe original invoice has already been paid, your credit note will be sent to your accounts payable department, and is credited back against the PO used for the purchase and put back into your budget. If your budget has been closed and this is no longer possible, you can request a refund by check instead at us.credits@abcam.com. The credit can also serve as a reference for a discount on a future purchase. I hope you have found this information helpful. Please do not hesitate to contact us with any additional questions you may have.

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Answer

Thank you for taking time to provide this information and for contacting us. I am sorry to hear this antibody is not providing satisfactory results and that these suggestions did not help.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I wish to apologize for the inconvenience. If you have purchase this item within the last six months I am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Answer

Thank you for contacting us.

The expected molecular weight of this protein is 549KDa. As recommended by our https://www.abcam.com/index.html?pageconfig=resource&rid=11403#B1 the bigger the size of the protein of interest, the lower the percentage of gel. I would suggest using at least an 8% gel (as used by another user in the https://www.abcam.com/index.html?datasheet=20737&tab=abreviews&intabreviewid=12715 which gave excellent results) and using reduced and denaturing conditions.

I hope this helps. Please do not hesitate to contact us for further assistance.

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1-10 of 26 Abreviews or Q&A

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