Overview

  • Product name
    Anti-ATF-4 antibody - C-terminal
    See all ATF-4 primary antibodies
  • Description
    Goat polyclonal to ATF-4 - C-terminal
  • Host species
    Goat
  • Specificity
    This antibody is expected to recognize both reported isoforms (NP_001666.2; NP_877962.1).
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human, Pig
    Predicted to work with: Mouse, Rat, Cow
  • Immunogen

    Synthetic peptide corresponding to Human ATF-4 aa 339-351 (C terminal) (Cysteine residue).
    Sequence:

    C-EEVRKARGKKRVP


    Database link: P18848
    (Peptide available as ab22854)

  • Positive control
    • WB: K562 cell lysate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: 0.05% Tris, 0.5% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab1371 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
WB Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 38 kDa).Can be blocked with Human ATF-4 peptide (ab22854).

Target

  • Function
    Transcriptional activator. Binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. It binds to a Tax-responsive enhancer element in the long terminal repeat of HTLV-I.
  • Sequence similarities
    Belongs to the bZIP family.
    Contains 1 bZIP domain.
  • Cellular localization
    Cytoplasm. Cell membrane. Nucleus. Colocalizes with GABBR1 in hippocampal neuron dendritic membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • Activating transcription factor 4 antibody
    • ATF 4 antibody
    • ATF4 antibody
    • ATF4 protein antibody
    • ATF4_HUMAN antibody
    • cAMP-dependent transcription factor ATF-4 antibody
    • cAMP-responsive element-binding protein 2 antibody
    • CREB 2 antibody
    • CREB-2 antibody
    • CREB2 antibody
    • Cyclic AMP dependent transcription factor ATF 4 antibody
    • Cyclic AMP response element binding protein 2 antibody
    • Cyclic AMP-dependent transcription factor ATF-4 antibody
    • Cyclic AMP-responsive element-binding protein 2 antibody
    • DNA binding protein TAXREB67 antibody
    • DNA-binding protein TAXREB67 antibody
    • Tax Responsive Enhancer Element B67 antibody
    • Tax-responsive enhancer element-binding protein 67 antibody
    • TaxREB67 antibody
    • TXREB antibody
    see all

Images

  • Anti-ATF-4 antibody - C-terminal (ab1371) at 0.3 µg/ml + K562 cell lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Huang T  et al. Curcumin mitigates axonal injury and neuronal cell apoptosis through the PERK/Nrf2 signaling pathway following diffuse axonal injury. Neuroreport 29:661-677 (2018). Read more (PubMed: 29570500) »
  • Xia X  et al. Arginine Supplementation Recovered the IFN-?-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2a Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells. Mol Cells 39:410-7 (2016). WB . Read more (PubMed: 27025389) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (vascular smooth muscle cells)
Permeabilization
Yes - 0.1 % triton X-100
Specification
vascular smooth muscle cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Methanol:Acetone 1:1

Abcam user community

Verified customer

Submitted Dec 29 2015

Question
Answer

Reviewing purchases of this product, the lab purchased ab1371 and not ab50546. Furthermore, in the methods section of the paper an anti-goat secondary antibody is mentioned but no goat primary is stated as being used. Therefore we do believe the researchers did use ab1371 but incorrectly cited this in the paper.

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Question
Answer

Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Je me permets de vous transférer le numéro de commande du remplacement gratuit de ce produit : xxxxxx. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (kidney)
Loading amount
30 µg
Specification
kidney
Gel Running Conditions
Reduced Denaturing (4-12 %)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 15 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3)
Loading amount
50 µg
Specification
3T3
Treatment
Cells transfected with plasmid encoding ATF4 with HA tag
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Aug 12 2011

Question

Hello,thank you for your reply. There was a second question that you did not touch on.I need to buy positive controls for both antibodies namelly C32 nuclear lysate and Raji cell lysate. Could you please e mail me cataloug numbers for them and if possible soon because I would like to place an order today. Thank you In response to your enquiry (ref: 760868):I have trouble using P 21 and ATF-4 antibodies(from other supplieres) in a western blot of mouse heart protein extracts. All literature mentiones cell extracts. I would like to order these 2 antibodies from your company. What is your experience using them on tissue extracts in Western blots?Where can I get the appropriate positive controls listed on data sheets? We have an answer Thank you for your enquiry.I am pleased that you are considering purchasing our p21 and ATF-4 antibodies. With regards the application of these antibodies using tissue lysates; p21 antibody (ab7960) have been applied by Xiaobo Chen using human smooth muscle cells. He has left a favourable review as have many of our customers who have applied this antibody using western blot against whole cell lysates. It has been shown to recognize murine p21 in a broad range of applications.ATF4 antibody (ab1371) has been shown to work by western blotting. It is predicted to cross react with the murine homolog based on sequence homology.I hope this information helps, please do not hesitate to contact us if you need any more advice or information.You can return to the datasheet for the product by clicking on:https://www.abcam.com/go.cfmp=4&ab=7960&v=52365

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Answer

Thank you for getting back to me. I am sorry that I did not provide you with reference to the positive controls for each of these antibodies. The following positive control should be used for ATF4 antibody (ab1371): Raji cell lysate: ab7908 Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate The following positive controls are recommended for use with p21 antibody (ab7960): C32 nuclear lysate or Colon carcinoma. Unfortunately we do not sell either of these as lysates. I am sorry I cannot be of more assistance on this part. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your phone call and I apologize for the shortage that you received. I have arranged for a free of charge replacement vial to be sent to you. It is being shipped to your attention on order reference # 99647 and you should receive this tomorrow. Please let me know if you have any additional questions or concerns.

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Question

BATCH NUMBER 63832 ORDER NUMBER 49259 DESCRIPTION OF THE PROBLEM Non-specific band. In addition to a high background (we are using the Abcam anti-goat secondary antibody raised in donkey) we do not see any band of the expeted size. We have several bands as it seems to see the case for other users. The most intense of them are approx. at 100 Kda, 60 Kda and 55 Kda...but anything at all between 40-45. SAMPLE We are using whole cell extracts from rat C6 glioma PRIMARY ANTIBODY We tried two different dilutions of primary antibody 1:500 and 1:200 in TBS-t 0.1%. We also peformed three different conditions for incubation with 1ry antibody in three different membranes: 1 h room temperature, 4?C o/n and 4?C o/n plus 1 h room temperature. Washes after primary antibody incubation: 2 quick washes, 1x10' and 2x5' with TBSt 0.1 % SECONDARY ANTIBODY We used the abcam anti-goat 2ry atibody raised in donkey (1:2000) in TBSt DETECTION METHOD Home made ECL (we regularly eploy it in WB) SAMPLE PREPARATION We employ the standard Lysis buffer with the cocktail with protease inhibitors (aprotin,leupeptin, etc) and also microcsytin 1 uM, PMSF and beta mercaptoethanol AMOUNT OF PROTEIN LOADED We loaded around 35 micrograms of protein per well ELECTROPHORESIS/GEL CONDITIONS (PAGE-SDS) 10-15 % acrylamide TRANSFER AND BLOCKING CONDITIONS transfer: 1,5 h at 100 v blocking 1 h with TBS plus 0.1% tween and 5% dried deffated milk HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Time of incubation and dilution of the primary antibody

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Answer

I'm sorry to hear you've had trouble with ab1371. Thank you for taking the time to fill in the self-help section to contact us, the information you have given me is very useful. I think you have taken all the right steps to get the antibody working and I will arrange a refund for this antibody immediately. As you mentioned, a number of customers have reported similar results in their preparations and we will investigate this further. Once again my sincere apologies,

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Answer

Thank you for your enquiry and providing more details of your assay. As we understand from your e-mail, the main problem you are experiencing is that you have a lot of non-specific bands. We have done our Western blot analysis with rabbit anti-goat IgG HRP and have only one distinct band at around 40 kDa (using Raji cell lysate). As we have indicated in our previous e-mail, we have not tested this primary antibody in hamster so we do not now if it works in this species. Reading through your protocol we see that you use 5% non-fat dry milk for blocking and a bovine anti-goat IgG conjugated with HRP as secondary antibody. Since the blocking agent is milk and the secondary antibody was raised in bovine, it is probable that there is a cross-reaction. The other alternative is that cow (bovine) and goat - these two species - phylogenetically are not very different from each other and the secondary antibody cross-reacting with endogenous immunoglobulins in the sample For more information about how to choose an antibody, please take a look at the following website: https://www.abcam.com/index.html?pageconfig=tech&mode=choosing Therefore we would suggest testing the secondary antibody alone (run a “secondary control” without the primary antibody) to see if you still have non-specific bands. We would also recommend using either a rabbit or a donkey polyclonal secondary antibody instead of bovine one. For your peace of mind, in case there may be some problem with the primary antibody, certainly we can offer you a replacement vial. If you would like to get it, then please do let us know and confirm your Purchase Order Number or Abcam Order Number. We are looking forward to hearing from you soon.

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Answer

Thank you for your message. As we understand from your e-mail you say that "hamster proteins are usually very similar to rat and mouse proteins". However, we would like to emphasize the fact that we haven't tested this particular antibody on hamster. It is possible that there is some cross-reaction with a 100 kDa protein in hamster that doesn't happen in human. All tested applications are specified on Abcam product datasheets. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.

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1-10 of 13 Abreviews or Q&A

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