Overview

  • Product name
  • Description
    Rabbit polyclonal to EIF2S1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human, Drosophila melanogaster
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human eIF2 alpha.

    Read Abcam's proprietary immunogen policy (Peptide available as ab26881.)

  • Positive control
    • Recombinant Human EIF2S1 protein (ab123468) can be used as a positive control in WB. HeLa Whole Cell Lysate Jurkat Whole Cell Lysate A431 Whole Cell Lysate NIH 3T3 Whole Cell Lysate MEF1 Whole Cell Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab26197 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 Use a concentration of 1 µg/ml.
IHC-P 1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).

Target

  • Function
    Functions in the early steps of protein synthesis by forming a ternary complex with GTP and initiator tRNA. This complex binds to a 40S ribosomal subunit, followed by mRNA binding to form a 43S preinitiation complex. Junction of the 60S ribosomal subunit to form the 80S initiation complex is preceded by hydrolysis of the GTP bound to eIF-2 and release of an eIF-2-GDP binary complex. In order for eIF-2 to recycle and catalyze another round of initiation, the GDP bound to eIF-2 must exchange with GTP by way of a reaction catalyzed by eIF-2B.
  • Sequence similarities
    Belongs to the eIF-2-alpha family.
    Contains 1 S1 motif domain.
  • Post-translational
    modifications
    Substrate for at least 4 kinases: EIF2AK1/HRI, EIF2AK2/PKR, EIF2AK3/PERK and EIF2AK4/GCN2. Phosphorylation stabilizes the eIF-2/GDP/eIF-2B complex and prevents GDP/GTP exchange reaction, thus impairing the recycling of eIF-2 between successive rounds of initiation and leading to global inhibition of translation (PubMed:15207627, PubMed:18032499). Phosphorylated; phosphorylation on Ser-52 by the EIF2AK4/GCN2 protein kinase occurs in response to amino acid starvation and UV irradiation.
  • Cellular localization
    Cytoplasmic granule. The cytoplasmic granules are stress granules which are a dense aggregation in the cytosol composed of proteins and RNAs that appear when the cell is under stress. Colocalizes with NANOS3 in the stress granules (By similarity).
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • EIF 2 alpha antibody
    • EIF 2 antibody
    • EIF 2A antibody
    • EIF 2alpha antibody
    • eIF-2-alpha antibody
    • eIF-2A antibody
    • EIF-2alpha antibody
    • EIF2 alpha antibody
    • EIF2 antibody
    • EIF2A antibody
    • EIF2S1 antibody
    • Eukaryotic translation initiation factor 2 subunit 1 alpha 35kDa antibody
    • Eukaryotic translation initiation factor 2 subunit 1 alpha antibody
    • Eukaryotic translation initiation factor 2 subunit 1 antibody
    • Eukaryotic translation initiation factor 2 subunit alpha antibody
    • IF2A_HUMAN antibody
    see all

Images

  • All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    ab26197 detects a band of ~ 36 kDa in HeLa, Jurkat and A431 whole cell lysates. This corresponds to the predicted molecular weight for eIF2 alpha according to Swiss prot. A number of other bands are detected at ~ 48, 23 and 15 kDa. We do not know the identity of these bands and we predict they represent non-specific binding.

  • All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 36 kDa

  • ICC/IF image of ab26197 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26197, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Image courtesy of Human Protein Atlas

    ab26197 staining eiF2 alpha in human small intestine. The paraffin embedded tissue was incubated with ab26197 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab26197 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray c

References

This product has been referenced in:
  • Takahashi H  et al. Identification of functional features of synthetic SINEUPs, antisense lncRNAs that specifically enhance protein translation. PLoS One 13:e0183229 (2018). Read more (PubMed: 29414979) »
  • Pareek G  et al. Loss of the Drosophila m-AAA mitochondrial protease paraplegin results in mitochondrial dysfunction, shortened lifespan, and neuronal and muscular degeneration. Cell Death Dis 9:304 (2018). Read more (PubMed: 29467464) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Pichia pastoris Cell lysate - whole cell (Pichia pastoris GS115 (his-) total lysate)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
4.4 µg
Specification
Pichia pastoris GS115 (his-) total lysate
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 06 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Arabidopsis thaliana Tissue lysate - whole (2-week-old seedlings)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
40 µg
Specification
2-week-old seedlings
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 Dec 03 2015

Answer

ab26197 is known to detect total protein levels of EIF2S1.

Read More

Answer

Thank you for your message and update.

I am sorry to hear the replacement vial has also not worked. I appreciate the time you have spent on these experiments and it is regrettable the results have not been successful.

As requested, I have asked our Finance department to issue a credit note for you.

Credit note ID: ####

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. Please do not hesitate to contact me if you have any further questions or concerns.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1153650.

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 6 month Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

With regards to ab5369, I am sorry we do not have any data to confirm if this antibody would be more sensitive. I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, anyEIF2S1 antibody tested in the species and application you are using should be suitable for your applications. Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done.

I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your reply.

I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and you prefer a credit note in this case, I will be pleased to arrange this for you.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

Here is thequestionnaire completed. Hope this could help to figure out what happens with this antibody.

Many thanks in advance. I am waiting your reply.

Sincerely,


Order Details
Antibody code: ab26197

Problem
Choose: Multiple bands and weak signal

Lot number: GR3277-1

General Information
Antibody storage conditions (temperature/reconstitution etc):-20ºC


Description of the problem (high background, wrong band size, more bands, no band etc.): Multiple bands and the right band size is weaker then all others.


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.): protein extract


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.): protein extract was obtained from mice tumors by using the RIPA buffer with protease and phosphatase inhibitors.


Amount of protein loaded: ˜100ug


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.): 12% SDSdenaturating gel


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.): Transfer in 1x TGM 1h @100v. membrane was blocked 3hours in 5%BSA in TBS-T.


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): ab26197 antibody was diluted in 5%BSA in TBS-T to the concentration of 1.25ug/ul. Incubation was performed o/n @ 4ºC, afterwards membrane was washed 4x 5min @ RT with TBS-T



Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): the secondary antibody was the goat anti-rabbit IRDye 680 from LI-COR Biosciences (cat# 926-32221) at 1/10000 dilutionin5%BSA in TBS-T. Incubation was performed 2h @ RT and membrane was washed 3x 5 min in TBS-T and the final wash step was done 15 min in TBS at RT


Detection method (ECL, ECLPlus etc.): Odissey


Positive and negative controls used (please specify): none



Optimization attempts (problem solving): I tried more time of incubation and I loaded more sample.
How many times have you tried the Western? 3 times


Have you run a "No Primary" control?No

Do you obtain the same results every time?No
e.g. are the background bands always in the same place?


What steps have you altered?

Additional Notes: When I loaded more sample and increased the incubation time I got the 3 bands showed in the picture because the first two attempts I didn't get anything.

Read More
Answer

Thank you for taking the time to complete our questionnaire. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

1. I would appreciate if you are able to provide an image which would help us to assess the results.

2. We recommend to load 20 - 30 ug of protein per lane of the gel to ensure it is not overloaded.

3. Could you confirm if the transfer to the membrane and quality of the sample assessed using a loading control?

4. I can recommend to try some fresh samples.

5. Could you confirm if thecurrent vial of secondary antibody working well with other primary antibodies? The concentration of secondary antibody may need to be reduced in order to help optimize the results.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and in mouse, human and fruit fly samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to annotate the molecular weight markers in the image, and describe what is in each lane.This would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details

Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (thorax extracts)
Loading amount
30 µg
Specification
thorax extracts
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Kiyoung Kim

Verified customer

Submitted May 04 2012

1-10 of 11 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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