Overview

  • Product name
  • Description
    Rabbit polyclonal to DAZL
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Chimpanzee, Common marmoset
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 to the C-terminus of Mouse DAZL.

    Read Abcam's proprietary immunogen policy (Peptide available as ab27733.)

  • Positive control
    • Testis (Human) Whole Tissue Lysate and Testis (Mouse) Whole Tissue Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab34139 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 at an assay dependent concentration. PubMed: 22156371
IHC-P Use a concentration of 0.5 - 5 µg/ml.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).

Target

  • Function
    RNA-binding protein, which is essential for gametogenesis. Plays a central role during spermatogenesis. May act by binding to the 3'-UTR of mRNA and thereby regulating the translation of key transcripts.
  • Tissue specificity
    Testis specific.
  • Sequence similarities
    Belongs to the RRM DAZ family.
    Contains 1 DAZ-like domain.
    Contains 1 RRM (RNA recognition motif) domain.
  • Domain
    The DAZ-like domain mediates the interaction with DAZAP1 and DAZAP2.
  • Cellular localization
    Cytoplasm. Nucleus. Predominantly cytoplasmic (By similarity). Nuclear in spermatogonia until near the end of the meiotic prophase and cytoplasmic localization from then onward.
  • Information by UniProt
  • Database links
  • Alternative names
    • DAZ homolog antibody
    • DAZ like autosomal antibody
    • DAZ-like autosomal antibody
    • DAZH antibody
    • DAZL 1 antibody
    • DAZL antibody
    • DAZL_HUMAN antibody
    • DAZL1 antibody
    • DAZLA antibody
    • Deleted in azoospermia like 1 antibody
    • Deleted in azoospermia like antibody
    • Deleted in azoospermia like autosomal antibody
    • Deleted in azoospermia-like 1 antibody
    • Deleted in azoospermia-like antibody
    • Germline specific RNA binding protein antibody
    • MGC26406 antibody
    • Spermatogenesis gene on the Y like autosomal antibody
    • SPGY like autosomal antibody
    • SPGY-like-autosomal antibody
    • SPGYLA antibody
    • Tpx2 antibody
    see all

Images

  • ab34139 staining DAZL in Mouse testis tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 3% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (1/1000) for 1 hour at 20°C. Ab150081 (1/500) was used as the secondary antibody. Phalloidin (red) was used as a counterstain staining F-actin.

    See Abreview

  • IHC image of DAZL staining in mouse testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab34139, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ab34139 at 1/100 staining human blastocyst cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 18 hours. An Alexa-Fluor ® 488 conjugated secondary antibody was used.

    See Abreview

  • The image, submitted as part of an Abreview, shows DAZL staining (green) of a human blastocyst. The antibody stains the cytoplasm of cells of the inner cell mass and trophoblast. DAZL is known to be located to the cytoplasm and DAZL transcripts have previously been discovered in the inner cell mass and trophblast (see PMID: 15879466).
  • The image shows staining of human testis tissue. The cytoplasm of Leydig cells and other cells in the seminiferous tubules stained positive. The antigen retrieval techniques used were Citrate at pH6 or Tris EDTA at pH9.
  • All lanes : Anti-DAZL antibody (ab34139) at 1 µg/ml

    Lane 1 : Human testis tissue lysate - total protein (ab30257)
    Lane 2 : Testis (Mouse) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 130 kDa, 70 kDa, 95 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 2 minutes
  • Ab34139 staining DAZL on juvenile marmoset testes by immunohistochemistry. The tissue was paraformaldehyde fixed and blocked in 1% milk for 1 hour at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 9 hours at 4°C in 5% milk. An Alexa Fluor 555 conjugated goat anti-rabbit antibody was used as the secondary. Nuclear staining shown in blue with DAPI.

    See Abreview

  • Anti-DAZL antibody (ab34139) at 1 µg/ml + GST tagged Recombinant Human DAZL protein (ab112279) at 0.1 µg

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 33 kDa


    Exposure time: 10 seconds
  • IHC-Fr image of DAZL (ab34139) staining mouse embryonic day 13 gonads. The sections were fixed in Paraformaldehyde and permeabilized in 0.3% Triton, before blocking in 1% BSA solution for 1 hour at 25°C. The primary antibody was added at a 1:100 dilution, and then incubated for 12 hours at 4oC. Anti-Dazl antibody (red) shows specific cytoplasmic staining only in primordial germ cells (Oct4, green), and not in surrounding somatic cells (DAPI, blue).

    See Abreview

References

This product has been referenced in:
See all 43 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
IHC - Wholemount
Sample
Quail Embryo (Stage HH11 Primordial Germ Cells)
Specification
Stage HH11 Primordial Germ Cells

Mr. David Huss

Verified customer

Submitted Jul 05 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Mar 30 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/sodium citrate
Sample
Mouse Tissue sections (adult testes)
Specification
adult testes
Permeabilization
No
Fixative
4%PFA

Dr. Qing Wen

Verified customer

Submitted May 06 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Mouse Tissue sections (adult testes)
Specification
adult testes
Permeabilization
No
Fixative
4%PFA

Dr. Qing Wen

Verified customer

Submitted May 01 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: antigen unmasking solution
Sample
Mouse Tissue sections (Testis)
Specification
Testis
Permeabilization
No
Fixative
Paraformaldehyde

Mr. Dan Stummer

Verified customer

Submitted Apr 09 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Sample
Chimpanzee Tissue sections (adult testis)
Specification
adult testis
Permeabilization
No
Fixative
Formaldehyde

Dr. 煜欽 林

Verified customer

Submitted Nov 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic day 13 gonads)
Specification
Embryonic day 13 gonads
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Marmoset (common) Tissue sections (Juvenile testes, spermatogonia cell population)
Specification
Juvenile testes, spermatogonia cell population
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Antigen retrieval solution by Wako / 105 degree @ 5 mins
Permeabilization
No
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Dr. 煜欽 林

Verified customer

Submitted Jul 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (Testis)
Specification
Testis
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer (pH 6)
Permeabilization
Yes - Triton X
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: RT°C

Dr. Daniel Murta

Verified customer

Submitted Sep 21 2011

Answer

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab34139. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support but please bear in mind that this product has not been tested using frozen sections (IHC-Fr). I have looked at the protocols you used and have a few questions and suggestions that might help you determine the cause of the no specific staining results. I would therefore appreciate if you can please clarify the following items. Since the cellular localization of this protein is predominantly cytoplasmic (nuclear in spermatogonia until near the end of the meiotic prophase and cytoplasmic localization from then onwards), you will need to fix your samples with ice cold methanol/acetone (1-10min), which also serves to permeabilize you samples for easy detection. For protocol details, please refer to out website protocol page. From my experience, samples prepared for frozen sections do not require overnight fixation (can be done for shorter time if desired). Usually, samples are embedded in OCT right after dissection. The reason is because usage of PFA can cause masking of the epitope and will require antigen retrieval methods. This maybe why you are not detecting any signals. Can you confirm that the secondary alone (no-primary) control you performed, did not show any signals? Or did you get background? Did the secondary antibody also managed to produce good results with other primary antibodies? In some cases, the problem may be with the secondary antibody not working properly. If the above suggestions did not help to improve your results, then I am afraid that this product is indeed not suitable for use in IHC-Fr application. In any case, can I encourage you to submit an Abreview via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to other researchers. We reward each Abreview with Abpoints which can be used for discounts off future purchases and gifts. If you decide to submit an Abreview, please include a note that you have already spoken to me, so that I will be able to quickly publish your Abreview, as I have already tried troubleshooting with you. To find out more about our Abreview system, please see the following URL: https://www.abcam.com/abreviews Should you require any further information or assistance, please do not hesitate to contact me.

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1-10 of 11 Abreviews or Q&A

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