Overview

  • Product name
  • Description
    Rabbit polyclonal to Stella
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Mouse Stella.

    Read Abcam's proprietary immunogen policy (Peptide available as ab23324.)

  • Positive control
    • This antibody gave a positive signal for Western Blot in Mouse Embryonic Germ Cell, Embryonic Stem Cell and Embryonic Carcinoma Cell Lysates. This antibody gave a positive signal for Immunofluorescence in Mouse Embryonic Stem cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab19878 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-Fr Use a concentration of 4 µg/ml.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function
    May play a role in maintaining cell pluripotentiality.
  • Tissue specificity
    Low expression in testis, ovary and thymus. Expressed in embryonic stem and carcinoma cells. Highly expressed in testicular germ cell tumors.
  • Developmental stage
    Expressed in fetal ovary.
  • Cellular localization
    Nucleus. Cytoplasm.
  • Information by UniProt
  • Database links
  • Form
    Stella is a gene specifically expressed in primordial germ cells, oocytes, preimplantation embryos and pluripotent cells. It encodes a protein with a SAP-like domain and a splicing factor motif-like structure, suggesting possible roles in chromosomal organization or RNA processing. Embryos without Stella are compromised in preimplantation development and rarely reach the blastocyst stage.
  • Alternative names
    • Compaction associated protein 1 antibody
    • Developmental pluripotency associated 3 antibody
    • Developmental pluripotency associated protein 3 antibody
    • Developmental pluripotency-associated protein 3 antibody
    • Dppa3 antibody
    • DPPA3_HUMAN antibody
    • PGC7 antibody
    • Stella related protein antibody
    • Stella-related protein antibody
    see all

Images

  • ICC/IF image of ab19878 stained mouse embryonic stem 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 (ab19878, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.
  • The image shows ab19878 staining of a crysosection of mouse embryonic genital ridges (E14.5). The samples were fixed overnight in 4% paraformaldehyde, permeabilised with 0.1% Triton and stained overnight at 4 degrees. 4µg/ml of antibody was used. Staining was in the nucleus and cytoplasm of Oct4-positive cells. The blue fluorescence is DAPI staining of DNA.

  • All lanes : Anti-Stella antibody (ab19878) at 1 µg/ml

    Lane 1 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate
    Lane 2 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
    Lane 3 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate
    Lane 4 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate with Mouse Stella peptide (ab23324) at 2 µg/ml
    Lane 5 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate with Mouse Stella peptide (ab23324) at 2 µg/ml
    Lane 6 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate with Mouse Stella peptide (ab23324) at 2 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 17 kDa
    Observed band size: 23 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 125 kDa, 25 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • ICC/IF image of ab19878 stained mouse embryonic stem cells. The cells were 4% formaldehyde 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 (ab19878, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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) at a concentration of 1.43µM.

  • Immunofluorescence analysis of porcine induced pluripotent stem cells cultured in 2i + LIF medium, staining Stella with ab19878.

    Cells were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked in 5% BSA. Cells were incubated with primary antibody (1/60) overnight at 4°C.

References

This product has been referenced in:
  • Leopardo NP & Vitullo AD Early embryonic development and spatiotemporal localization of mammalian primordial germ cell-associated proteins in the basal rodent Lagostomus maximus. Sci Rep 7:594 (2017). IHC-P ; Other species . Read more (PubMed: 28377629) »
  • Wang H  et al. Induction of Germ Cell-like Cells from Porcine Induced Pluripotent Stem Cells. Sci Rep 6:27256 (2016). ICC/IF ; Pig . Read more (PubMed: 27264660) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Germline stem cells)
Permeabilization
Yes - 1% Triton X in PBS
Specification
Germline stem cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 21 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, embryonic day 15.5)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, embryonic day 15.5
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Apr 06 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Loading amount
200000 cells
Specification
mouse embryonic stem cell
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Oct 24 2012

Answer

Thank you for contacting us.

Your credit note ID is XXXXX.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Answer

Thank you for contacting us. I am sorry to hear 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. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (embryonic stem cell)
Specification
embryonic stem cell
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Oct 11 2012

Question

1) Abcam product code ab19878
2) Abcam order reference number or product batch number GR77041-1
3) Description of the problem Nuclear staining in multiple tissues (such as skeletal muscle, epithelium of the small intestine, cartilage, brown adipose tissue, spongiotrophoblasts, and others) that should be negative for DPPA3; cytoplasmic staining in pancreas; no staining in heart muscle. This pattern was found consistently in 4 experiments on E18 and E19 mouse fetuses and placentas.
4) Sample preparation:
Species
Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: Carnoy's fixed mouse fetuses and placentas on E18 and E19 of gestation.
Sample preparation 7 µm paraffin sections
Positive control Fetal gonads
Negative control Rest of fetus
5) Fixation step
Yes/No
If yes: Fixative agent and concentration Carnoy's fixative (60% ethanol, 30% CHCl3, 10% acetic acid)
Fixation time over night (10-12 h)
Fixation temperature 4 °C
6) Antigen retrieval method Citrate buffer, pH 6.0, 10 min at 480 W in microwave, cooling period 40-50 min
7) Permeabilization method: None
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration:
8) Blocking agent (eg BSA, serum…): Dako antibody dilution solution (S3022)
Concentration
Blocking time 30 min
Blocking temperature RT
9) Endogenous peroxidases blocked? 3.5% H2O2 in PBS, 10 min @ RT
Endogenous biotins blocked? No
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution Between 1 : 200 to 1 : 400 (recommended by abcam 1:250)
Diluent buffer Dako antibody dilution medium (S3022)
Incubation time
11) Secondary antibody:
Species: Swine (DAKO P0399)
Reacts against: Rabbit
Concentration or dilution 1 : 100
Diluent buffer Dako antibody dilution medium (S3022)
Incubation time 60 min at RT
Fluorochrome or enzyme conjugate HRP
12) Washing after primary and secondary antibodies:
Buffer 1 x PBS
Number of washes 3 x 5 min
13) Detection method HRP
14) How many times have you run this staining? 4 x
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody? Different concentrations of primary AB
Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem. Picures show
a-DPPA3 1/200 E19 cartl = nuclear anti-DPPA3 staining in cartilage of E19 mouse fetus (dilution of primary AB 1 : 200)
a-DPPA3 1/400 E19 cartl = nuclear anti-DPPA3 staining in cartilage of E19 mouse fetus (dilution of primary AB 1 : 400) (dilution of primary AB 1 : 400)
a-DPPA3 1/400 E19 intest = nuclear anti-DPPA3 staining in small intestine of E19 mouse fetus; staining is stronger in epithelial cell nuclei towards the tip of the villus!
(dilution of primary AB 1 : 400)
DPPA3 contr wo 1st E19 cartl = no primary antibody, concentration of 2ndary AB as in the other pictures

Read More
Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided; the protocol looks fine to me to however I have following further suggestions for improvements;

- Please try Tris EDTA buffer or enzymatic antigen retrieval method; it may well be that heating tissue for 10 minutes have damaged the tissue which is why antibody is binding all over.
- Please try blocking in 10% non immune swine serum or 5% BSA for at least 2 hours.
- Ethanol or methanol fixation is recommended for frozen sections; the antigen retrieval method can skipped as Ethanol/methanol helps unmasking the antigen for antibody binding. Please try one section without antigen retrieval step.

The rest of the protocol is fine. Please let me know how the staining go.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult ovary)
Specification
Adult ovary
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. 煜欽 林

Verified customer

Submitted Aug 29 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Marmoset (common) Tissue sections (Marmoset adult testis)
Specification
Marmoset adult testis
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Target retrieval solution by Dako
Permeabilization
Yes - Triton-X
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%

Dr. 煜欽 林

Verified customer

Submitted Aug 09 2012

1-10 of 12 Abreviews or Q&A

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