Overview

  • Product name
    Anti-hCG beta antibody [5H4-E2]
    See all hCG beta primary antibodies
  • Description
    Mouse monoclonal [5H4-E2] to hCG beta
  • Host species
    Mouse
  • Specificity
    The antibody recognizes the whole hCG beta and the C-terminal end part of hCG beta, as well as the hCG dimer molecule. It does not react with hCG alpha, FSH, LH, TSH and albumins (evaluated by simple-binding ELISA using 1.00 µg/well purified antigens (WHO standards) and 1.00 µg/ml primary antibody concentrations). This antibody labels the cytoplasm of hCG beta producing (trophoblastic) cells in placenta, mola hydatidosa, choriocarcinoma, embryonal carcinoma and some undifferenciated lung cancers, as well as, the JAR choriocarcinoma cell line. It does not react with cells of normal endometrial, testicular tissues and lung (tested by immunohistochemistry on formalin-fixed, paraffin-embedded tissue sections using the Streptavidin-biotinylated peroxidase method).
  • Tested applications
    Suitable for: WB, ELISA, IHC-P, IHC-Fr, ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Horse
  • Immunogen

    Full length native protein (purified) corresponding to Human hCG beta.
    Database link: P01233

  • Positive control
    • ICC/IF: Human chorionic villus cells. IHC-P: Human fetal chorionic villus tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab9582 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.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. It gives optimal immunostaining with the majority of well-known detection systems (e.g. LSAB, ABC/SABC, PAP, APAAP, double staining, etc.). Antigen retrieval, such as heating of slides in 10 mM citrate buffer, pH 6.00 or proteolytic pre-treatment of tissues may increase the staining intensity. The recommended time of incubation is 60 minutes at room temperature.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

Images

  • Formalin-fixed, paraffin-embedded human seminoma tissue stained for hCG beta with ab9582 at a 1/50 dilution in immunohistochemical analysis.

  • ab9582 at 1/100 dilution staining hCG beta in human chorionic villus cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde, without permeabilization and blocked in 3% BSA for 1 hour at 25°C. The primary antibody was diluted in PBS (3% BSA) and incubated with sample at 4°C for 12 hours. An Alexa Fluor® 568 conjugated Goat polyclonal to mouse IgG was used at 1/500 dilution as secondary. Image A show DAPI staining, B show staining with ab9582 and C is the merged image which show staining with DAPI and ab9582 together. 

    See Abreview

  • ab9582 staining hCG beta in human fetal chorionic villus by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Samples were fixed in 4% paraformaldehyde and then incubated with ab9582 in PBT [3% bovine serum albumin (BSA) and 0.5% Triton X-100 in phosphate-buffered saline (PBS)] overnight at 4°C. After extensive washing with PBT, samples were incubated with a secondary antibody conjugated to Alexa-Fluor® 488 for 1 hour at room temperature in the dark. After nuclear staining with Hoechst slides were mounted with PBS/ glycerol.

References

This product has been referenced in:
  • Sengodan SK  et al. BRCA1 regulation on ß-hCG: a mechanism for tumorigenicity in BRCA1 defective breast cancer. Oncogenesis 6:e376 (2017). Read more (PubMed: 28869585) »
  • Kolahi K  et al. Real-Time Tracking of BODIPY-C12 Long-Chain Fatty Acid in Human Term Placenta Reveals Unique Lipid Dynamics in Cytotrophoblast Cells. PLoS One 11:e0153522 (2016). IF . Read more (PubMed: 27124483) »
See all 6 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for contacting Abcam. Included in this email are the instructions for returning your item/s to Abcam. When you return the goods, please ensure that you follow the procedures as below: - Return in the original Abcam box with the cooler box and ice pack (or similar) within 7 days from today. - Return all documents (or copies of) that arrived with the product (if they’re available – should be invoice, delivery note and datasheet) - Please return via an overnight courier such as UPS, DHL, or FEDEX. - Ship to the following address: Please be advised that a 20% restock fee will apply to this order. The most up-to-date information regarding your order, invoice, and credit note status is available online in your Abcam account information. If your order was placed by credit card, your refund will be issued to your credit card. If your order was placed by Purchase Order, all customers must submit a credit note with their invoice to their Accounts Payable department to be handled. The Accounts Payable must have a copy of the credit note so the original invoice can be updated to reflect the correct amount owed. If you have any other questions please do not hesitate to contact us! Email: US.orders@Abcam.com Phone: 888-77-ABCAM(22226) Fax: 866 739 9884 Customer Service hours of operation: Monday-Friday 8:30 AM- 9:00PM EST

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Answer

Thank you for contacting Abcam regarding ab9582.


I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

I regret to inform to that we have not tested it in our lab so I am afraid that we can't advise on the binding characteristics to PDMS (Polydimethylsiloxane) surface. As I have mentioned in my previous e-mail, ab9582 has not been tested in immunoprecipitation.

Apologies!

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Answer

Thank you for your enquiry and your interest in our products.

I can confirm that ab9582 (Anti-hCG beta antibody [5H4-E2]), has not been tested for IP yet, though the isotype has been determined (IgG1).

Normally, we would suggest using Protein G for mouse IgG1 antibodies; whilst for moue IgG2a and mouse IgG2b can be used with both Protein A and G. Please could you advise your customer to take a look at this site for further information:

https://www.abcam.com/index.html?pageconfig=resource&rid=11385

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for your enquiries.

I can confirm that ab9582hCG beta antibody was raised against the beta subunit ofhCG only, and its reactivity was blocked by betasubunit of hCG, notthe alpha subunit. Therefore, we would not expect it to detect the alpha form.

Human chorionic gonadotropin is a heterodimeric protein, with an alpha and betasubunit:

The alpha subunit is 92 amino acids long and isidentical tothe alpha subunit ofluteinizing hormone (LH), follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH).

The beta subunit is unique to hCG.It contains 145 amino acids, encoded by six highly-homologous genes that are arranged in tandem and inverted pairs on chromosome 19q13.3 - CGB (1, 2, 3, 5, 7, 8).

There is information on the beta subunit of hCG protein on the UniProt protein database page from the link on the datasheet:

http://www.uniprot.org/uniprot/P01233

There is information on the apha subunit on the following page:

http://www.uniprot.org/uniprot/P01215

I hope this will be helpful. I suggest the customer may like to considera further literature search in order to obtain further information.

If you have any further questions, please do not hesitate to contact me.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Chorionic villus)
Specification
Chorionic villus
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Paola Spitalieri

Verified customer

Submitted Jan 19 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Horse Recombinant protein (culture broth)
Loading amount
2 µg
Specification
culture broth
Blocking step
Milk as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 5

Dr. Anna Ubach

Verified customer

Submitted Jun 19 2006

Answer

Thank you for your feedback regarding ab9582. You mention a band at a molecular weight similar to that of albumin, is this around 60-70KDa? I think the MW for albumin is about 66KDa. Could this band be explained by the fact the antibody recognizes the whole hCG beta and the C-terminal end part of hCG beta as well as the hCG dimer molecule? I believe the molecular weight of a monomer of hCG is about 38K, which is about half this weight. Please do not hesitate to contact me should you need further information,

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Answer

Thank you for the information that you have provided. We have not had any other complaints concerning this antibody. Since you have not been able to obtain a signal with the new vial under the same conditions that you used with the previous lot, I would like to send you a replacement vial to try. It's a possibility that this latest vial went off during the shipping process. However, I do need either the Abcam order number or purchase order number that was used to order ab9582. Thank you, and I look forward to hearing from you.

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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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