Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732)

Overview

  • Product nameAnti-Melanoma antibody [HMB45 + DT101 + BC199]
    See all Melanoma primary antibodies
  • Description
    Mouse monoclonal [HMB45 + DT101 + BC199] to Melanoma
  • Tested applicationsSuitable for: Flow Cyt, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    HMB45 - Pigmented melanoma metastases from LN MART-1 - Recombinant human MART-1 protein

  • EpitopeHMB45 and MART-1
  • Positive control
    • Metastatic melanoma in lymph node.
  • General notesMelanoma Cocktail is HMB45[HMB45] + two different clones of MART-1[M2-7C10] + [M2-9E3]. HMB34 isotype: IgG1/kappa and both clones of MART-1: IgG2b + IgG2b


    HMB-45 and MART-1 are coexpressed in the majority of melanomas, as well as uniquely expressed in certain cases. Thus, the HMB-45 and MART-1 cocktail is potentially more sensitive than HMB-45 or MART-1 alone. MART-1 is a cocktail of clones DT101 and BC199. The combination of HMB-45 and the MART-1 cocktail make this triple antibody cocktail a first-order pan melanoma screener.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: Proprietary, not sodium azide or thimerosal.
    Constituents: PBS, Tissue Culture Supernatant. pH 7.3; Protein carrier.
  • Concentration information loading...
  • Primary antibody notesHMB-45 and MART-1 are coexpressed in the majority of melanomas, as well as uniquely expressed in certain cases. Thus, the HMB-45 and MART-1 cocktail is potentially more sensitive than HMB-45 or MART-1 alone. MART-1 is a cocktail of clones DT101 and BC199. The combination of HMB-45 and the MART-1 cocktail make this triple antibody cocktail a first-order pan melanoma screener.
  • ClonalityMonoclonal
  • Clone numberHMB45 + DT101 + BC199
  • Myelomaunknown
  • IsotypeIgG2b
  • Light chain typekappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab732 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
Flow Cyt Use at an assay dependent concentration. PubMed: 22147606
IHC-P Use at an assay dependent concentration. The staining intensity of this melanoma cocktail is enhanced by performing heat mediated antigen retrieval using a high pH buffer. The HMB-45 portion of this antibody will stain with citrate buffer, pH 6.0; however, data shows the MART-1 portion of this cocktail may be negative unless a high pH buffer is used. We therefore, strongly recommend that you do not use citrate buffer for this antibody cocktail.
IHC-Fr Use at an assay dependent concentration.
For an ABC system, dilute 1/50-1/100 and incubate for 30-60 minutes at RT. It is sometimes difficult to interpret DAB stained melanomas due to endogenous pigment, therefore we recommend you substitute an AEC or a Fast Red substrate protocol.

Target

  • RelevanceMalignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre existing benign nevus, which occurs most often in the skin but also may involve other sites.
  • Cellular localizationMembrane; Single-pass membrane protein
  • Database links
  • Alternative names
    • Antigen LB39 AA antibody
    • Antigen SK29 AA antibody
    • MART1 antibody
    • melan A antibody
    • Melan A protein antibody
    • Melanoma antigen recognized by T-cells 1 antibody
    • Monophenol monooxygenase antibody
    • Tumor rejection antigen AB antibody
    • tyrosinase antibody
    see all

Anti-Melanoma antibody [HMB45 + DT101 + BC199] images

  • ab732 staining murine melanoma metastasis tissue sections by IHC-P.  Tissue sections were paraformaldehyde fixed and blocked with 0.5% Perkin-Elmer TNB Blocking Buffer for 30 minutes at 25°C.  The primary antibody was diluted 1/50 and incubated for 18 hours at 4°C.  An Alexa Fluor 488® conjugated goat anti-mouse was used as the secondary

    Confocal image shows melanoma in magenta, vessels in red (Collagen Type IV, ab19808), and melanin in cyan. Tissue is perfusion-fixed 15µm cryostat sections mounted on slides (i.e., lightly fixed, but not paraffin embedded).

    See Abreview

References for Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732)

This product has been referenced in:
  • Turajlic S  et al. Whole-genome sequencing reveals complex mechanisms of intrinsic resistance to BRAF inhibition. Ann Oncol 25:959-67 (2014). ICC/IF ; Human . Read more (PubMed: 24504448) »
  • Girotti MR  et al. Inhibiting EGF receptor or SRC family kinase signaling overcomes BRAF inhibitor resistance in melanoma. Cancer Discov 3:158-67 (2013). IHC ; Human . Read more (PubMed: 23242808) »

See all 9 Publications for this product

Product Wall

Thank you for contacting Abcam.
I have attached the Abcam, mouse on mouse staining protocol to see if that can give you any suggestions. If you can send me a copy of the protocol you are using, then I would be happy to see if there are any suggest...

Read More

I am sorry to confirm we do not currently have fluorochrome conjugated Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) on our catalog.


However, you may like to consider either using a secondary antibody (http://www.abcam.com/index....

Read More

Our Melanoma Cocktail is HMB45[HMB45] + two different clones of MART-1[M2-7C10] + [M2-9E3].
HMB34 isotype: IgG1/kappa and both clones of MART-1: IgG2b + IgG2b
Epitope/antigen: HMB45 and MART-1

Thank you very much for contacting us with your enquiry.
I am sorry to hear that there has been some variability with ab732. We don't have any other recent reports of the antibody not working, and there haven't been any changes to the antibody, so...

Read More

Thank you for your enquiry. I am pleased to provide the following suggested positive controls for these antibodies. I have mainly chosen tissue that has been tested succesfully and shown in an image on the datasheets: 1.Anti-ErbB 2 antibody (ab2428) Hu...

Read More

Thank you for your patience while I was looking into this. I have heard back from the lab and they have told me that ab732 was tested using the DIVA decloaking agent (http://biocare.net/wp-content/uploads/DV2004X.pdf) which is at pH.6.2. Also for the r...

Read More

Thank you for contacting Abcam. The 3 clones in the ab732 cocktail do recognize mouse tissue. The IHCP protocol that was used to test the antibody was optimized to human tissue and the following protocol was used: Pretreatment Protocol Heat Retrieval M...

Read More
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 (murine melanoma metastasis)
Specification murine melanoma metastasis
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Blocking step (agent) for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 07 2008

Thank you for your enquiry. You would not want to use the citrate buffer for antigen retrieval on your samples when using this antibody cocktail. You can use 1mM EDTA pH 8.0, and you should determine the optimal heating time for your sample. A goo...

Read More

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