Overview

  • Product name
    Anti-HOXB13 antibody [mAbcam53931]
    See all HOXB13 primary antibodies
  • Description
    Mouse monoclonal [mAbcam53931] to HOXB13
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human HOXB13.

    Read Abcam's proprietary immunogen policy (Peptide available as ab57024.)

  • Positive control
    • This antibody gave a positive signal against the HOXB13 (Human) Recombinant Protein.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab53931 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-P Use at an assay dependent concentration. PubMed: 25745994
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 57 kDa corresponding to the tagged recombinant HOXB13 protein
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Sequence-specific transcription factor which is part of a developmental regulatory system that provides cells with specific positional identities on the anterior-posterior axis.
  • Sequence similarities
    Belongs to the Abd-B homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Homeo box B13 antibody
    • Homeobox B13 antibody
    • Homeobox protein Hox B13 antibody
    • Homeobox protein Hox-B13 antibody
    • Homeobox protein HoxB13 antibody
    • HOX B13 antibody
    • HOXB 13 antibody
    • HOXB13 antibody
    • HXB13_HUMAN antibody
    • PSGD antibody
    • PSGD protein antibody
    see all

Images

  • Anti-HOXB13 antibody [mAbcam53931] (ab53931) at 5 µg/ml + HOXB13 Recombinant protein at 0.1 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 31 kDa
    Observed band size: 57 kDa
    why is the actual band size different from the predicted?



    ab53931 recognizes the tagged HOXB13 recombinant protein which has an expected molecular weight of 57 kDa.
  • ICC/IF image of ab53931 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53931, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HepG2 cells fixed in 4%PFA at 1ug/ml..
  • Overlay histogram showing HeLa cells stained with ab53931 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab53931, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Jiang Y  et al. Repression of Hox genes by LMP1 in nasopharyngeal carcinoma and modulation of glycolytic pathway genes by HoxC8. Oncogene 34:6079-91 (2015). IHC-P ; Human . Read more (PubMed: 25745994) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

The antibody will detect a band at ˜37 kDa, the image given on the datasheet show 57 kDa which is becuase the GST tagged recombinant protein was used.

I hope this new info will also be helpful.

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Question

A customer had problems from ab53931. As follows:

客户姓名 殷雪东 单位 重庆医科大学附属第一医院 联系电话 023-89011477 货号 ab53931 lot 819179 order no 953260
1) Abcam product code ab53931
2) Abcam order reference number or product batch number :lot:819179
3) Storage conditions: -20℃
4) Description of the problem (high background, no band, wrong band size...)
No band
Predicted band size : 57
Observed band size : two bands:95 and 110
5) Sample preparation:
Name of the tissue/cell line: normal breast tissues
Sample species: human
Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates
Lysis buffer : RIPA lysis buffer
Protease inhibitors: cocktail protease inhibitor
Phosphatase inhibitors:
Reducing agent ?
Boiling for ≥5 min? yes
Protein loaded ug/lane or cells/lane 50ug
Positive control
Negative control
6) Percentage of gel 10%
Type of membrane PVDF
Protein transfer verified: YES
Blocking agent and concentration :5% BSA
Blocking time 1 hour
Blocking temperature room temp
7) Primary antibody (If more than one was used, describe in “additional notes”) : Rabbit monoclonal to HOXB13.
Concentration or dilution 1:400
Diluent buffer:5% BSA
Incubation time : over l night
Incubation temperature: 4℃
8) Secondary antibody: 中衫金桥
Species: goat
Reacts against: rabbit
Concentration or dilution 1:1000
Diluent buffer: 5% BSA
Incubation time: 1 hour
Incubation temperature: room temp
Fluorochrome or enzyme conjugate: HRP
9) Washing after primary and secondary antibodies: TBST
Buffer
Number of washes three times
10)Detection method:chemical exposure
11) How many times have you run this staining? Tiwce
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody? NO, we use the standard protocol which can be successfully used for other antibodies.

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Answer

Thank you very much for contacting us. I am sorry to hear that you are experiencing problems with the antibody.

The protocol looks fine to me however I would like to draw your attention to the following details

- ab53931 is Mouse monoclonla antibody. You will need an anti mouse secondary antibody.

Please try the experiment with correct secondary antibody and let us know the results.

This product is out of Abpromise guarantee because it was purchased in January 2011. We however are happy in providing troubleshooting suggestions, which will help in getting the right band size.

I hope this info will be helpful. Should you have any other question please do not hesitate to ask.

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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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