Overview

  • Product name

  • Description

    Rabbit polyclonal to ZC3H13
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rabbit, Horse, Cow, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide corresponding to a region between residues 1175 and 1225 of Human ZC3H13 (NP_055885.2)

  • Positive control

    • Whole cell lysate from HeLa, 293T and NIH3T3 cells.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituents: 1.815% Tris, 1.764% Sodium citrate, 0.021% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab70802 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
ICC/IF 1/400.
IP Use at 2 µg/mg of lysate.
WB 1/2000 - 1/10000. Detects a band of approximately 240 kDa (predicted molecular weight: 197 kDa).

Target

  • Sequence similarities

    Contains 1 C3H1-type zinc finger.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Information by UniProt
  • Database links

  • Alternative names

    • 2600010B19Rik antibody
    • 3110050K21Rik antibody
    • 4930570G11Rik antibody
    • C87618 antibody
    • DKFZp434D1812 antibody
    • FLJ35669 antibody
    • KIAA0853 antibody
    • ZC3H13 antibody
    • ZC3HD_HUMAN antibody
    • Zinc finger CCCH domain-containing protein 13 antibody
    • zinc finger CCCH type containing 13 antibody
    see all

Images

  • ab70802 (1/400) staining ZC3H13 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% TritonX100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • All lanes : Anti-ZC3H13 antibody (ab70802) at 0.04 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg
    Lane 5 : NIH3T3 whole cell lysate at 50 µg

    Developed using the ECL technique.

    Predicted band size: 197 kDa
    Observed band size: 240 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 200 kDa, 85 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • Detection of ZC3H13 by Western Blot of Immunoprecipitate.
    ab70802 at 1µg/ml staining ZC3H13 in HeLa whole cell lysate previously immunoprecipitated using ab70802 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane).
    Detection: chemiluminescence with exposure time of 30 seconds.

References

This product has been referenced in:

  • Huang B  et al. Cyclophosphamide Regulates N6-Methyladenosine and m6A RNA Enzyme Levels in Human Granulosa Cells and in Ovaries of a Premature Ovarian Aging Mouse Model. Front Endocrinol (Lausanne) 10:415 (2019). Read more (PubMed: 31316467) »
  • Knuckles P  et al. Zc3h13/Flacc is required for adenosine methylation by bridging the mRNA-binding factor Rbm15/Spenito to the m6A machinery component Wtap/Fl(2)d. Genes Dev 32:415-429 (2018). Read more (PubMed: 29535189) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your reply.

The concentration of ab70802 is 1 mg/ml. Yes, the recommended antibody dilution is 1/2000 - 1/10000. But the antibody can be used at other dilutions as well, dependenton the samples being studied.

As secondary antibody any HRP-conjugated anti-rabbit secondary antibody can be used, such as e.g.ab16284 (https://www.abcam.com/Rabbit-IgG-secondary-antibody-ab16284.html) or ab97051 (https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-HRP-ab97051.html).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question
Answer

Thank you for contacting us.

The following WB protocol has been used with this antibody, togetherwitha 3-8% SDS Precast gel and HeLa whole cell lysate.

Western Blot Protocol

Reagents Needed:

20X Running Buffer


Tricine (free base) 71.7 g
Tris (free base) 72.6 g
SDS 10.0 g
Sodium Bisulfite 2.5 g



Adjust to 500 ml with ultra pure water.

Store at 4°C. For 1X Running Buffer, add 10 ml of 20X Running Buffer to 190 ml of distilled water.

10X Transfer Buffer


Tris (free base) 15.2 g
Glycine 72.1 g
SDS 5.0 g



Ultra pure water to 500 ml

Store at 4°C.

1X Transfer Buffer


10X Transfer Buffer 50 ml
Methanol 100 ml
Distilled water 350 ml



Make fresh for each use.

5% non-fat dry milk in TBST


Carnation non-fat dry milk 50 g
TBST 1 liter



TBST (Tris Buffered Saline with Tween 20, pH8.0)


Tris 6.1 g
NaC l 8.68 g
Tween-20 500 mcl



Adjust the volume to 1 liter with distilled water. Adjust pH to 8.0 with HCL.

Store at 4-25°C.

Electrophoresis in SDS-Polyacrylamide Gel and Transfer to Nitrocellulose:

Molecular weight markers:ab116027, ab116028, ab115832, ab119210.
Nitrocellulose membranes.

Cut open the package that contains the gel cassette and drain away the buffer.
Rinse the wells with distilled water.
Rinse the wells with fresh 1x running buffer.
Place the gels on the buffer core so that the shorter plates face inward. If only using one gel, use a buffer dam to seal the other side.
Fill the inner core with fresh 1X running buffer. If there are no leaks, fill the outer core with running buffer. Load samples.
Run the gels at 150V until the dye front reaches the bottom of the gel (approximately 60 minutes).
Soak nitrocellulose membrane and blotting paper in 1X transfer buffer for at least 5 minutes prior to opening gel cassette.
Open gel cassettes and place the gel on the nitrocellulose membrane sandwiched between two pieces of blotting paper.
Place in transfer apparatus and fill with fresh 1X transfer buffer.
Run transfer apparatus for 60-75 minutes on 35V.



Western Blotting:

Remove the membrane from the transfer apparatus and place in 20 ml of 5% non-fat dry milk in TBST for one hour, with gentle shaking.
Dilute the primary antibody in 15 ml of 5% non-fat dry milk in TBST. For best results, the optimal dilution of antibody should be empirically defined.
Incubate the membrane in diluted primary antibody for two hours to overnight with gentle shaking at room temperature.
Wash the membrane three times, 10 minutes each time in TBST.
Dilute the secondary HRP conjugated antibody in 15 ml of reconstituted 5% non-fat dry milk in TBST. For best results, the optimal concentration of the secondary HRP conjugated antibody should be empirically defined.
Incubate the membrane in diluted anti-Rabbit IgG-HRP Conjugate for 60 minutes.
Wash as directed in step 4.
Develop blots with substrate solution and place in plastic membrane protector.
Expose membrane to film or CCD camera.




I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TritonX100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Nov 17 2009

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