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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hCG beta antibody [5H4-E2] (ab9582)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hCG beta antibody [5H4-E2] (ab9582)

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

  • Immunocytochemistry/ Immunofluorescence - Anti-hCG beta antibody [5H4-E2] (ab9582)
    Immunocytochemistry/ Immunofluorescence - Anti-hCG beta antibody [5H4-E2] (ab9582)This image is courtesy of an Abreview submitted by Paola Spitalieri.

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hCG beta antibody [5H4-E2] (ab9582)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hCG beta antibody [5H4-E2] (ab9582)Image from Spitalieri P et al, Cloning Stem Cells. 2009 Dec;11(4):535-56, Fig 1, DOI: 10.1089=clo.2009.0046.

    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

Publishing research using ab9582? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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9 reviews or Q&A

10
Votes: Highest First

Also, we recently purchased an antibody from ABCAM (#ab9582) together with an Elisa kit, but it turns out we don’t need the antibody anymore. We received it last Friday, Oct26, and is still in the closed package, but I would like to know if it’s possib...

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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...

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I am using this ab in frozen sections of human placenta and obtaining no signal.
PFA fixed tissues
Block: Serum free block or milk contaiing 0.1% Triton-100, 30 min at RT
1' ab - no primary, 1:50, 1:100, 1:200
2' ab - tested both polyme...

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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 th...

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Thanks for your kindly reply, this customer also wants to know whether protein A or protein G can coat on the PDMS (Polydimethylsiloxane) surface and binding with the ab9582 (Anti-hCG beta antibody [5H4-E2]) or not?

Please help this customer...

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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 immunoprecip...

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Dear technical support team: This customer raises an enquiry about ab7399 (Protein A protein) and ab84187 (Protein A protein), she wants to know which one is more suitable to bind with ab9582 (Anti-hCG beta antibody [5H4-E2]), could you please help thi...

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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 usi...

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Thanks for your kindly reply, this customer wants conduct ICC/IF with JAR cell and she also wants to know what’s different between alpha and beta form hCG, could you please help this customer to solve the problem?

Thanks for your kindl...

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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 ...

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Immunocytochemistry/ Immunofluorescence abreview for Anti-hCG beta antibody [5H4-E2]

 Good
Abreviews
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
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abreview image
Username

Paola Spitalieri

Verified customer

Submitted Jan 19 2010

Western blot abreview for Anti-hCG beta antibody [5H4-E2]

 Poor
Abreviews
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
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Username

Dr. Anna Ubach

Verified customer

Submitted Jun 19 2006

We have encounter in our test with the new Ab a huge cross-reactivity with a protein exhibiting the same electrophoretic mobility than albumin. Therefore, the cross-reactivity with the specific protein hCG is considerable small. Let me know if you can...

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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 th...

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Essentially our problem is that this antibody (ab9582) worked very well when we purchased approximately 2 years ago, in at least 3 independent experiments. Recently (one month ago) we purchased more of the same antibody and now we can not reproduce the...

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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 woul...

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