Overview

  • Product name
  • Description
    Guinea pig polyclonal to PDX1
  • Host species
    Guinea pig
  • Tested applications
    Suitable for: IHC-FoFr, IHC-Fr, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fusion protein containing N-terminal sequence from mouse PDX1

Properties

Applications

Our Abpromise guarantee covers the use of ab47308 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-FoFr Use at an assay dependent concentration. PubMed: 20170518
IHC-Fr 1/200.
IHC-P Use at an assay dependent dilution.
ICC/IF 1/10000.

Target

  • Function
    Activates insulin, somatostatin, glucokinase, islet amyloid polypeptide and glucose transporter type 2 gene transcription. Particularly involved in glucose-dependent regulation of insulin gene transcription. Binds preferentially the DNA motif 5'-[CT]TAAT[TG]-3'. During development, specifies the early pancreatic epithelium, permitting its proliferation, branching and subsequent differentiation. At adult stage, required for maintaining the hormone-producing phenotype of the beta-cell.
  • Tissue specificity
    Duodenum and pancreas (Langerhans islet beta cells and small subsets of endocrine non-beta-cells, at low levels in acinar cells).
  • Involvement in disease
    Defects in PDX1 are a cause of pancreatic agenesis (PAC) [MIM:260370]. This autosomal recessive disorder is characterized by absence or hypoplasia of pancreas, leading to early-onset insulin-dependent diabetes mellitus. This was found in a frameshift mutation that produces a truncated protein and results in a second initiation that produces a second protein that act as a dominant negative mutant.
    Defects in PDX1 are a cause of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853]; also known as diabetes mellitus type 2. NIDDM is characterized by an autosomal dominant mode of inheritance, onset during adulthood and insulin resistance.
    Defects in PDX1 are the cause of maturity-onset diabetes of the young type 4 (MODY4) [MIM:606392]; also symbolized MODY-4. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
  • Sequence similarities
    Belongs to the Antp homeobox family. IPF1/XlHbox-8 subfamily.
    Contains 1 homeobox DNA-binding domain.
  • Domain
    The Antp-type hexapeptide mediates heterodimerization with PBX on a regulatory element of the somatostatin promoter.
    The homeodomain, which contains the nuclear localization signal, not only mediates DNA-binding, but also acts as a protein-protein interaction domain for TCF3(E47), NEUROD1 and HMG-I(Y).
  • Post-translational
    modifications
    Phosphorylated by the SAPK2 pathway at high intracellular glucose concentration.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Glucose sensitive factor antibody
    • Glucose-sensitive factor antibody
    • GSF antibody
    • IDX 1 antibody
    • IDX-1 antibody
    • IDX1 antibody
    • Insulin promoter factor 1 antibody
    • Insulin promoter factor 1 homeodomain transcription factor antibody
    • Insulin upstream factor 1 antibody
    • IPF 1 antibody
    • IPF-1 antibody
    • IPF1 antibody
    • Islet/duodenum homeobox 1 antibody
    • Islet/duodenum homeobox-1 antibody
    • IUF 1 antibody
    • IUF-1 antibody
    • IUF1 antibody
    • MODY4 antibody
    • Pancreas/duodenum homeobox 1 antibody
    • Pancreas/duodenum homeobox protein 1 antibody
    • pancreatic and duodenal homeobox P antibody
    • PDX 1 antibody
    • PDX-1 antibody
    • PDX1 antibody
    • PDX1_HUMAN antibody
    • Somatostatin transactivating factor 1 antibody
    • Somatostatin-transactivating factor 1 antibody
    • STF 1 antibody
    • STF-1 antibody
    • STF1 antibody
    see all

Images

  • Immunohistochemical analysis of developing murine pancreas taken from wild-type (+/+) and SEL1L-null mice (-/-). PDX1 (green) was stained with ab47308.

    Tissue was fixed with paraformaldehyde before freezing and sectioning. Sections were permeabilized with 0.2% Triton X-100 for 20 min and blocked with 5% normal donkey serum and 1% BSA in PBS at room temperature for at least 1 hour. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) overnight at 4°C. A Cy2®-conjugated donkey anti-guinea pig IgG (1/500) was used as the secondary antibody.
  • Immunohistochemical analysis of paraformaldehyde-fixed,0.1 % Triton X-100 permeabilized mouse pancreas tissue labelling PDX1 with ab47308 at 1/200 dilution,followed by Goat Anti-guinea pig IgG conjugation Alexa Fluor®488 secondary antibody at 1/200 dilution.

    See Abreview

  • Immunostaining of islets from 4 week-old mice with ab47308 guinea pig anti-PDX1 (red) and an anti-insulin antibody (green).

References

This product has been referenced in:
  • Memon B  et al. Enhanced differentiation of human pluripotent stem cells into pancreatic progenitors co-expressing PDX1 and NKX6.1. Stem Cell Res Ther 9:15 (2018). IHC-P ; Human . Read more (PubMed: 29361979) »
  • Aigha II  et al. Differentiation of human pluripotent stem cells into two distinct NKX6.1 populations of pancreatic progenitors. Stem Cell Res Ther 9:83 (2018). Read more (PubMed: 29615106) »
See all 13 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6.0
Sample
Mouse Tissue sections (pancreas)
Specification
pancreas
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH6.0
Sample
Human Tissue sections (pancreas)
Specification
pancreas
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2013

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (8 % gel)
Sample
Human Cell lysate - whole cell (Pancreatic progenitors from hPSCs)
Specification
Pancreatic progenitors from hPSCs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 10 2013

Question

Please find the answers to the questionnaire.
Images are attached.
I still hope that the technical teams are able to provide me solutions,
otherwise I'd like to have refund for this product.
IHC Questionnaire
It will take approximately 5 minutes to complete the questionnaire.
Please fill-in all fields so that we can better assist you
1) Abcam product code ab 47308
2) Abcam order reference number 1081728 or product batch number
3) Description of the problem false positive staining
4) Sample preparation: pancreas
Species human
Type of sample: Fresh frozen sections, perfusion fixed frozen sections,
PFA/formalin fixed paraffin embedded sections, cells in culture, other:
Sample preparation
Positive control N/T
Negative control normal IgG
5) Fixation step
Yes/No
If yes: Fixative agent and concentration 4% Paraformaldehyde (for frozen
sections)
Fixation time 15min
Fixation temperature room temperature
6) Antigen retrieval method for FFPE sections, xylene dewax, followed by
either sodium citrate buffer (pH6.0) for 10min or 25min / Tris-EDTA (pH9.0)
for 20min / EDTA (pH8.0) for 20min in scientific microwave
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing
agent in any dilution buffers? Yes
Permeabilizing agent and concentration: Two methods were tested
1st method: diluting primary antibodies in dilution buffer (1xPBS, 0.1%
Triton-X100) and incubate for 2hrs at R.T.
2nd method: before incubating with primary antibody, sections were
permeabilized using 1xPBS buffer, 2% Triton X, 0.2% Tween20 for 1hr.
8) Blocking agent (eg BSA, serum…): normal goat serum as secondary
antibody was anti-guinea pig raised in goat
Concentration 3%
Blocking time blocked simultaneously with primary antibody incubation
(for 2hrs)
Blocking temperature room temperature
9) Endogenous peroxidases blocked?
Endogenous biotins blocked?
10) Primary antibody (If more than one was used, describe in "additional
notes") :
Concentration or dilution 1:50
Diluent buffer 1xPBS, 0.1% TritonX-100
Incubation time 2hrs at r.t.
11) Secondary antibody: Fluorescent Texas Red
Species: goat
Reacts against: guinea pig IgG
Concentration or dilution 1:200
Diluent buffer 1xPBS, 0.1% TritonX-100
Incubation time 1hr
Fluorochrome or enzyme conjugate Texas Red
12) Washing after primary and secondary antibodies:
Buffer 1xPBS
Number of washes 3times
13) Detection method fluorescent microscope 40x magnification
14) How many times have you run this staining? 3
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody?
Antigen retrieval buffer, permeabilization
Document attachment: Attaching images of your IHC is strongly recommended
and can greatly speed up our investigation of your problem.

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, I would like to offer some suggestions to help optimize the results from ab47308 Anti-PDX1 antibody:
1. As you are performing a heat- induced antigen retrieval, an extra permeabilization step is not necessary for the FFPE.
2. Triton can be quite harsh on some epitopes, therefore, you may increase the clarity of the staining by reducing the Triton concentration during . Still, I can recommend keeping the Triton to your incubation solution to facilitate the antibodies solubility, but in a concentration of 0.025%.
3. As the necessary permeabilization step for the frozen sections, I would like to suggest to decrease the permeabilization step time with 0.1% Triton-X100in PBS down to 30 min.
4. To assure that the non-specific binding site are blocked, please block with at least 5% serum for 30 min before the antibody incubation.
5. The antibody concentration is very high compared to what we suggest on the datasheet, especially as the used tissue is the positive control. I would therefore suggest to increase the specificity of the antibody by decreasing the concentration down to at least 1:500 and to incubate overnight at 4C.
Hopefully, this will bring down the non-specific binding. Should the suggestions not improve the results, please do let me know.
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, and I thank you for your cooperation.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Pancreas)
Specification
Pancreas
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 16 2012

Answer

Thank you for contacting us.

PDX1 will be expressed in the nucleus ofpancreatic islet and intercalated duct cellsin adult mice. I hope this helps, please let me know if you need any additional information.

Read More

Answer

 I have issued a free of charge replacement with the order number ****. This is for rabbit anti-PDX1, ab47267, to replace ab47308. The fixation is with 4% PFA for 5 minutes, followed by permeabilization with Triton X-100. The full protocol is below. Staining Cell cultures. 1. Rinse cells with PBS. 2. Fix in 4% paraformaldehyde in PBS for 5 minutes. 3. Wash 3 times with cold PBS for 5 minutes. 4. Permeablilize membrane with 0.2% Triton X for 10 minutes. 5. Wash twice with cold PBS for 5 minutes. 6. Block with 10% goat serum in PBS for 30 minutes. 7. Add primary antibody (1/5000) for 4 hours. 8. Wash 3 times for 10 minutes. 9. Add secondary goat anti-rabbit antibody (Alexa546) at 1/1000 for 1 hour. 10. Wash with PBS for 5 minutes. 11. wash with DAPI (1/10 000) for 10 minutes. 12. Wash with PBS for 10 minutes. 13. Mount with fluorescent mounting media. 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.  

Read More

Answer

I assume you have the antibody ab47267, as that is the only rabbit anti-PDX1 in our catalogue that has been tested for ICC. We do not have samples of any of our antibodies but I can send a full vial as a replacement, perhaps from a lot different from what you have already tried. If the fresh vial gives a poor result again, we can send a different antibody if you like. Do you know the catalogue number of the rabbit PDX1 antibody that you have, and the lot number?

Read More

Answer

I am sorry to read that this is still not working for you. My guess is that the fixation is still too long at 2 hours, even with 4% vs. 10% PFA, or that there is lot-to-lot variability with regards to compatibility with formaldeyhe-based fixation. If you like, I will replace the antibody but I am concerned you will have the same result if I send ab47308. Are you interested in trying a different PDX1 antibody? We do not have another antibody raised in guinea pig but we have one from goat, ab47383, and another from rabbit, ab47267. If you do not want to try one of these (or any other primary antibody in our catalogue) free-of-charge, I can issue a credit or refund. I look forward to your reply.

Read More

Answer

Following up my previous message, the laboratory and I think the problem may be the 10% PFA fixation for 10 minutes at room temperature. The lab used 4% PFA and fixed for 2 hours at 4C. I think the fixation time could be made shorter than two hours, for instance 30 minutes at 4C. This will preserve the structure of the cell, but will hopefully minimize the cross-linkages introduced by the PFA that may be hindering antibody:antigen recognition. The acetone/methanol approach is an alternative, which does not create chemical modifications of the PDX1, but the laboratory has not tried this. Your current fixation protocol may be fine for other antibody stains, but each antibody:antigen interaction is unique, and ab47308 may have difficulty with 10% PFA at RT. Please let me know how the antibody works for you if you try one of these fixation modifications, and please contact me if you have any questions.

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1-10 of 13 Abreviews or Q&A

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