Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab6028 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 1/500 - 1/1000. Detects a band of approximately 8 kDa (predicted molecular weight: 8 kDa).

Target

  • Function
    Could participate in the response to proapoptotic stimuli and promotes cellular growth in a way that helps the tissue counteract diverse injuries. May contribute to the metastatic phenotype.
  • Tissue specificity
    Highly expressed in pancreas.
  • Post-translational
    modifications
    Phosphorylated.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Candidate of metastasis 1 antibody
    • COM1 antibody
    • Nuclear protein 1 antibody
    • Nupr1 antibody
    • NUPR1_HUMAN antibody
    • p8 protein antibody
    • Protein p8 antibody
    see all

References

This product has been referenced in:
  • Palam LR  et al. Integrated stress response is critical for gemcitabine resistance in pancreatic ductal adenocarcinoma. Cell Death Dis 6:e1913 (2015). WB . Read more (PubMed: 26469962) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human colon cancer cell)
Loading amount
30 µg
Specification
human colon cancer cell
Gel Running Conditions
Reduced Denaturing (12.5% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 09 2013

Question

DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE HuCCT1, PanC1 whole cell lysates PRIMARY ANTIBODY Concentration or dilution : 1:1,000 Diluent buffer : 1% skim milk in PBST Incubation time : O/N Incubation temperature: at 4℃ wash Buffer : PBST Number of washes : 6 times(each for 5 min) DETECTION METHOD LAS-3000 POSITIVE AND NEGATIVE CONTROLS USED Positive control : PanC1 p8 has been shown to be highly expressed in PanC1 cells (PMID: 21344397) ANTIBODY STORAGE CONDITIONS Store at -20℃ SAMPLE PREPARATION Lysis buffer : RIPA buffer Protease inhibitors: 2mM PMSF and 1x PIC Phosphatase inhibitors : no Reducing agent : yes Boiling for ≥5 min? yes. AMOUNT OF PROTEIN LOADED Protein loaded ug : 50ug protein ELECTROPHORESIS/GEL CONDITIONS 20% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Type of membrane : PVDF Protein transfer verified : protein transfer was verified by dye transfer and commassie stainingof the gel Blocking agent and concentration : 1% skim milk in PBST Blocking time : 1 hour Blocking temperature : Room temperature SECONDARY ANTIBODY Jackson ImmunoResearch anti-Rabbit igG-HRP Concentration or dilution : 1:10,000 Diluent buffer : in PBST Incubation time : 2 hours Incubation temperature: room temperature wash Buffer : PBST Number of washes : 6 times(each for 5 min) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Blocking : 1% skim or 2% BSA Transfer : 1 hr or 40 min Primary Ab incubation : O/N or 3days ADDITIONAL NOTES Its predicted size is 8kDa but, some references showed 15~29kDa bands. There are non-specific bands only and any of them showed knock out pattern with siRNA treated sample.

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, it appears that ab6028 is not performing as well as might be expected. I would therefore like to offer you a free of charge replacement of this antibody with another from our catalogue. We have 4 other primary antibodies directed against p8: ab46889, ab93964, ab94034 and ab87454. All of which are suitable for WB. Please note ab87454 is a mouse monoclonal. Please let me know which antibody you would like to try and I will arrange it for you. Additionally, some suggestions which may help to optimise the results from ab6028 and the antibody which you choose to replace it with. -We would typically recommend blocking using 5% BSA solution -As the protein is small I would suggest removing the SDS from the transfer buffer if you haven't already as this hinders the binding to the membrane. I would also suggest including 20% methanol in the transfer buffer. -You may want to try reducing the transfer time as the protein is small it may migrate through the membrane too fast to be adsorbed efficiently More useful information may be found here: https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, if you have any further questions please don't hesitate to ask.

Read More

Question
Answer

Thank you for your enquiry. One difference that I see between these two antibodies is that ab6028 has more detail for the immunogen sequence "Synthetic peptide: ERKLVTKLQNSERKKRGARR, corresponding to amino acids 62-82 of Human p8" and ab6027 says that the immunogen is a synthetic peptide to the C terminus. Also, ab6027 has been used in a publication which is listed at the bottom of the data sheet. Other than the immunogen information, these antibodies appear to be identical. There is a price reduction on both of these antibodies until the 31st of December making ab6028 five dollars cheaper than ab6027. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Question

BATCH NUMBER 66531 ORDER NUMBER see end DESCRIPTION OF THE PROBLEM No signal at all, and the background is very clean (even after 30 minutes of exposure) SAMPLE 12 Human tumor cell lines including breast carcinoma (as positive control since the expression of p8 in breast cancer has been reported), prostate cancer, fibrosarcoma, cervix carcinoma and colorectal carcinoma. PRIMARY ANTIBODY abcam/rabbit anti-human/1:500/1 hour, 5 minutes 3 times in TBS-T SECONDARY ANTIBODY Oncogene/Goat anti-rabbit/1:3000/45 minutes, 5 minutes 3 times in TBS-T, then 5 minutes once in TBS DETECTION METHOD ECL, from Santa Cruz POSITIVE AND NEGATIVE CONTROLS USED Positive control for p8: 3 breast tumor cell line whole cell lysates ANTIBODY STORAGE CONDITIONS Performed one hybridization right after I received the antibody, i.e. before the aliquots and frozen. SAMPLE PREPARATION RIPA buffer with PMSF, Aprotinin and sodium orthovanadate. Samples were boiled for 5 minutes. AMOUNT OF PROTEIN LOADED approximately 30-50 ug/lane ELECTROPHORESIS/GEL CONDITIONS 15% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS 10x Blotting buffer: 1 L 30.3 g Trizma base (= 0.25 M) 144 g Glycine (= 1.92 M) pH should be 8.3; do not adjust. To make 2 L of 1x Blotting buffer: 400 ml Methanol 200 ml 10x Blotting buffer 1400 ml water transfered at 15V constant voltage overnight Blocking: Blotto A: 1x TBS, 5% milk, 0.05% Tween-20 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? increased the primary antibody concentration to 1:200 dilution ADDITIONAL NOTES the same blots were incubated with anti b-action primary antibody with same conditions, and positive signal were detected. the messenger RNA level are very high in some of the cell lines (the same as that of b-actin or even higher) according to my Real Time PCR results. my P.O. number does not fit in the field. It's UHN5-014546. The order was placed through Cedarlane.

Read More
Answer

I'm sorry to hear you are having problem with ab6028. P8 is a nuclear protein, do you do nuclear extraction to have a concentrated sample to load onto your gels? I would like to suggest an overnight incubation of the primary antibody (trying 1:200 and 1:500) at 4C diluted in TBST (0.1% Tween). I would also recommend human pancreatic cancer tissue as a positive control. Please let me know if this helps and do not hesitate to contact us again if you still have problems,

Read More

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

Sign up