Anti-GATA4 antibody (ab84593)

Rabbit polyclonal GATA4 antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Human, Rhesus monkey. Cited in 21 publication(s). Independently reviewed in 11 review(s).

Overview

  • Product name

  • Description

    Rabbit polyclonal to GATA4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Rhesus monkey
    Predicted to work with: Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human GATA4 aa 100-200 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab94555)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; HepG2; HEK293; Caco 2.

Properties

Applications

Our Abpromise guarantee covers the use of ab84593 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-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-Fr 1/50.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).

Target

  • Function

    Transcriptional activator that binds to the consensus sequence 5'-AGATAG-3' and plays a key role in cardiac development (PubMed:24000169). Involved in bone morphogenetic protein (BMP)-mediated induction of cardiac-specific gene expression (By similarity). Binds to BMP response element (BMPRE) DNA sequences within cardiac activating regions (By similarity). Acts as a transcriptional activator of ANF in cooperation with NKX2-5 (By similarity). Promotes cardiac myocyte enlargement (PubMed:20081228). Required during testicular development (PubMed:21220346). May play a role in sphingolipid signaling by regulating the expression of sphingosine-1-phosphate degrading enzyme, spingosine-1-phosphate lyase (PubMed:15734735).
  • Involvement in disease

    Atrial septal defect 2
    Ventricular septal defect 1
    Tetralogy of Fallot
    Atrioventricular septal defect 4
    Testicular anomalies with or without congenital heart disease
    GATA4 mutations can predispose to dilated cardiomyopathy (CMD), a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
  • Sequence similarities

    Contains 2 GATA-type zinc fingers.
  • Post-translational
    modifications

    Methylation at Lys-300 attenuates transcriptional activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • ASD2 antibody
    • GATA 4 antibody
    • GATA binding protein 4 antibody
    • GATA-binding factor 4 antibody
    • gata4 antibody
    • GATA4_HUMAN antibody
    • MGC126629 antibody
    • Transcription factor GATA 4 antibody
    • Transcription factor GATA-4 antibody
    • Transcription factor GATA4 antibody
    • VSD1 antibody
    see all

Images

  • All lanes : Anti-GATA4 antibody (ab84593) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 44 kDa
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • ab84593 staining GATA4 (green) in mouse embryo tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized in 1% Triton X-100 and blocked with 10% goat serum for 1 hour at room temperature; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50 in 10% goat serum) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody. Red - Myh6/7, Blue - DAPI.

    See Abreview

  • Immunofluorescence image of Gata4 (ab84593) on human embryonic stem cell derived cardiomyocytes. The cells were fixed in methanol and permeabilized in 0.1% Triton-X-100. In this image you can see a cardiomyocyte stained in red with a cardiac alpha actinin antibody and both the cardiomyocyte and a fibroblast stained in green with ab84593.

    See Abreview

  • All lanes : Anti-GATA4 antibody (ab84593) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Human Fetal Heart - Total Protein Tissue Lysate
    Lane 3 : Heart (Mouse) Tissue Lysate
    Lane 4 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 44 kDa
    Observed band size: 44 kDa
    Additional bands at: 110 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:

  • Anamthathmakula P  et al. Steroidogenic Factor 1 (Nr5a1) is Required for Sertoli Cell Survival Post Sex Determination. Sci Rep 9:4452 (2019). Read more (PubMed: 30872705) »
  • Vaez SA  et al. The cardiac niche role in cardiomyocyte differentiation of rat bone marrow-derived stromal cells: comparison between static and microfluidic cell culture methods. EXCLI J 17:762-774 (2018). Read more (PubMed: 30190666) »
See all 28 Publications for this product

Customer reviews and Q&As

1-7 of 7 Q&A

Answer

Thank you for contacting us. Your credit note ID is xxxxxx. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. As requested, I have also issued a free of charge replacement with the order number xxxxxx. To check the status of the order please contact our Customer Service team and reference this number. 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.

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Answer

Thank you for your reply. You do not need to return the antibodies which you have already received. Further we are very happy to send you new products against the same target or a different target. You do mentionAnti Neurofibromin ab17963, and I would be very happy to provide that for you. How would you like to proceed with the second antibody, new lot/vial, new product or credit note? I look forward to your reply. Please let me know if you have any questions.

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Answer

Thank you for your reply.

Based on the information that you have provided, I do not think there is anymore protocol advice that I could supply that would help resolve the issue you are having with these two antibodies in western blot. Therefore, I would be happy to replace or refund the cost of these antibodies for you.

Please let me know how you would like to proceed and I will be happy to help resolve this issue.

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Answer

Thank you for your reply and for providing that information.

For ab84593, the band that you are seeing at just above 50kDa, is very intense and I am wondering if that could be the correct band, but is just running a little higher than expected. What percentage of gel were you running; as maybe using a 12.5 or 15% gel would help resolve the band size a little more.

As for ab39688, you are correct that none of the bands that you are seeing would correspond to the correct molecular weight of the protein. Possibly incubating overnight at 4C, rather than at room temp for 2 hours, along with switching to a BSA block, may help to resolve the correct bands of interest.

Both of the antibodies are covered under our Abpromise for six months and are guaranteed to work in western blot on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

Please let me know, if changing the protocol conditions slightly do not improve the situation and I will be happy to replace or refund the antibodies for you.

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Question
Answer

I am sorry that you have been experiencing some technical issues with a few of our antibodies. Under our Abpromise, we guarantee our antibodies to work as stated on the datasheet. If they fail to work as stated to work as stated on the datasheet and we cannot determine why, then I am happy to replace or refund the cost of the antibody.

To be able to help you further with this, would you be able to provide a few protocol questions:

1 - What sample type are you using?

2 - Are you seeing any bands, no bands etc?

3 - How much sample are you loading?

4 - What type of blocking agent are you using?

5 - What primary antibody concentrations have you tried and how long do you incubate for?

We do take all complaints on products seriously and we do our best to solve these problems. I look forward to your reply and helping to resolve this issue.

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Question
Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unseres Wissens nach wurde keiner unserer anti GATA4 Antikörper bereits auf Hirsch getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von des bestellten Antikörpers abdecken, und gegen eine erneute Bestellung eines weiteren primären Antikörpers von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, welchen GATA 4 Antikörper Sie auf Hirsch testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in Hirsch.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderen primären Antikörpers zu dem gleichen Preis wie den bestellten anti GATA 4 Antikörper bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer vom GATA4 Antikörper bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Question

DESCRIPTION OF THE PROBLEM I am working with isolated day six chicken embryo hearts. When I probed my blot with GATA4 antibody purchased from abcam, I got several weak background-like bands and one strong band which is located at about 120KD, this is far away from the predicted size of 45KD or so. I understand that this antibody is designed for human, not chicken and I checked the antigen sequence, it is not completely identical but I'd say 70%-80% identical. Please see attached picture for the actual blot: please focus on the right side, the left one was detected with a different primary antibody. You can see the quite strong green band at about 120kd but that is way off the expected position. The third lane was messed by a bubble during transfer. Red blot is beta-actin which is 42KD. Is it possible to make this antibody work with chicken samples? Thank you. SAMPLE Chicken (Gallus gallus) Sample: Total protein extraction with RIPA buffer from isolated day six chicken embryo heart PRIMARY ANTIBODY Ab84593 at 1:1000 in TPBS, for one hour at room temperature. Washed four times with TPBS, each time for five minutes after detection. Also used sigma anti-betaactin(mouse) as loading control, which was put in at 1:10000. DETECTION METHOD Odyssey fluroscence detection. POSITIVE AND NEGATIVE CONTROLS USED None at this point. ANTIBODY STORAGE CONDITIONS Freezed at -20C aliquoted. No repeatly freeze and defreeze. SAMPLE PREPARATION Samples were homogenized in RIPA buffer with 1:100 protease inhibitor cocktail and 1mM DTT added, then shaked in cold room for two hours and centrifuged at 15000g, 10 min, the supernatant was used in blots. Laemilli buffer and NuPage reducing agent was used to prepare sample for western blot, heated at 70C for ten minutes before loading into wells. AMOUNT OF PROTEIN LOADED 25ug ELECTROPHORESIS/GEL CONDITIONS Reducing, bio-rad MiniProtean precast gels. 10%, tris-glycine buffer. TRANSFER AND BLOCKING CONDITIONS NuPage transfer buffer, wet transfer at 30V for one hour. Blocking agent was Rockland blocking buffer for near infrared western blot. SECONDARY ANTIBODY Odyssey's secondary: Goat anti mouse (for detection of beta-actin) and Goat anti rabbit (for detection of nkx2.5), both at 1:15000, incubatd at room temperature for one hour. Afterwards, four washes with TPBS, five minutes each were done.

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Answer

Thank you for contacting Abcam regarding ab84953.

I am sorry that you have been experiencing difficulties with this antibody in WB with your chicken samples. We have not tested chicken samples so this is not a species we guarantee, and as you mentioned, the antibody's immunogen does not share significant homology with the chick sequence, thus I am not sure you can expect cross reactivity.

I am not sure there are any protocol suggestions that will improve your results with this antibody. It seems that it is not suitable for chicken.

Please do not hesitate to contact us if you have any additional questions.

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