Overview

  • Product name

    Anti-eEF1A1+eEF1A2+eEF1AL3 antibody
  • Description

    Rabbit polyclonal to eEF1A1+eEF1A2+eEF1AL3
  • Host species

    Rabbit
  • Specificity

    Due to sequence homology the antibody recognises all three eEF1A forms, eEF1A1, eEF1A2 and eEF1A3. Through WB ab37969 was found to detect recombinant and endogenous eF1A1 and eF1A2.
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Rabbit, Horse, Chicken, Hamster, Cow, Cat, Dog, Pig, Chimpanzee, Macaque monkey
  • Immunogen

    Synthetic peptide corresponding to Human eEF1A1+eEF1A2+eEF1AL3 aa 200-300 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab37968)

  • Positive control

    • This antibody gave a positive signal in the following: Human Whole Cell Lysates (HeLa, Jurkat, A431, HEK 293, HepG2, MCF-7, SHSY-5Y, U2OS) Mouse Whole Cell Lysates (NIH 3T3, MEF1) Mouse Tissue Lysates (Brain, Liver, Heart, Kidney, Pancreas, Testis, Skeletal muscle, Spinal Cord, Ovary) Rat Whole Cell Lysate(PC12) Rat Tissue Lysate (Brain, Liver, Heart)

Properties

Applications

Our Abpromise guarantee covers the use of ab37969 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.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

Target

  • Relevance

    Function: This protein promotes the GTP-dependent binding of aminoacyl-tRNA to the A-site of ribosomes during protein biosynthesis. Tissue specificity eEF1A1: Brain, placenta, lung, liver, kidney, pancreas but barely detectable in heart and skeletal muscle. Tissue specificity eEF1A2: Brain, heart, and skeletal muscle. Similarity: Belongs to the GTP-binding elongation factor family. EF-Tu/EF-1A subfamily. PTM eEF1A1: ISGylated.
  • Database links

  • Alternative names

    • CCS3 antibody
    • eEF1A-1 antibody
    • eEF1A-2 antibody
    • EEF1A1 antibody
    • Eef1a2 antibody
    • EEF1AL antibody
    • EF 1 alpha 2 antibody
    • EF-1-alpha-1 antibody
    • EF-1-alpha-2 antibody
    • EF-Tu antibody
    • EF1A antibody
    • EF1A1_HUMAN antibody
    • EF1A2_HUMAN antibody
    • Elongation factor 1 A 2 antibody
    • Elongation factor 1 alpha antibody
    • Elongation factor 1 alpha 2 antibody
    • Elongation factor 1-alpha 1 antibody
    • Elongation factor 1-alpha 2 antibody
    • Elongation factor Tu antibody
    • Eukaryotic elongation factor 1 A-1 antibody
    • Eukaryotic elongation factor 1 A-2 antibody
    • Eukaryotic translation elongation factor 1 alpha 2 antibody
    • GRAF 1EF antibody
    • HS1 antibody
    • Leukocyte receptor cluster member 7 antibody
    • PTI1 antibody
    • Statin-S1 antibody
    see all

Images

  • All lanes : Anti-eEF1A1+eEF1A2+eEF1AL3 antibody (ab37969) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 whole cell lysate (ab7902)
    Lane 5 : HepG2 whole cell lysate (ab7900)
    Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : U2OS (Human osteosarcoma 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: 50 kDa

  • ICC/IF image of ab37969 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab37969, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal 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).

  • All lanes : Anti-eEF1A1+eEF1A2+eEF1AL3 antibody (ab37969) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 5 : Heart (Mouse) Tissue Lysate
    Lane 6 : Kidney (Mouse) Tissue Lysate
    Lane 7 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 8 : Testis (Mouse) Tissue Lysate
    Lane 9 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 10 : Spinal Cord (Mouse) Tissue Lysate
    Lane 11 : Ovary (Mouse) Tissue Lysate - normal tissue
    Lane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 13 : Brain (Rat) Tissue Lysate - normal tissue
    Lane 14 : Liver (Rat) Tissue Lysate
    Lane 15 : Heart (Rat) Tissue Lysate
    Lane 16 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

    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: 50 kDa
    Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.

  • All lanes : Anti-eEF1A1+eEF1A2+eEF1AL3 antibody (ab37969) at 1 µg/ml

    Lane 1 : Recombinant Human eEF1A1/EF-Tu protein (ab81792)
    Lane 2 : Human EEF1A2 full length protein (ab40317)

    Lysates/proteins at 0.1 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size: 50 kDa


    Exposure time: 3 minutes

References

This product has been referenced in:

  • Wu M  et al. URGCP/URG4 promotes apoptotic resistance in bladder cancer cells by activating NF-?B signaling. Oncotarget 6:30887-901 (2015). Read more (PubMed: 26429874) »
  • Dong C  et al. Proteasome modulates positive and negative translational regulators in long-term synaptic plasticity. J Neurosci 34:3171-82 (2014). Read more (PubMed: 24573276) »
See all 9 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Spinal Cord)
Permeabilization
No
Specification
Spinal Cord
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
Fixative
Paraformaldehyde

Herr Dr. Ralph Nawrotzki

Verified customer

Submitted Aug 17 2015

Answer

Thank you for your reply.

I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for you.

Credit note ID: 23754

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

Read More

Question

Dear Sir/Madam,

Thanks for your reply.

1.) I forgot to mention but I run my gel already under reducing conditions by adding 100mM DTT in my sample buffer when loading the samples on gel.

2.) The transfer of proteins worked out perfect as you can see on the image attached to this mail where you see the successful transfer of the other POI around 28-32 kDa.

3.) The dilutions I used were 1/1000 (as mentioned on the datasheet). A second time 1/500 and the last blot I used 1/250 for the ab37969. All this dilutions without success.

4.) The vial of secondary antibody I used for all my blots is perfectly working and I encountered never problems with my other rabbit antibodies.

I hope I've helped you with giving you this information.

At this moment the ab37969 antibodies still doesn't give me a proper working result, unfortunately.



Best regards,



Thomas Geuens



----------------------------------------------------------------------
Thomas Geuens, PhD Student
VIB - Department of Molecular Genetics
Peripheral Neuropathy Group
University of Antwerp - CDE
Parking P4, Building V, Room V1.17
Universiteitsplein 1
BE-2610 Antwerpen
Belgium

Tel: +32-3-265.10.27
Tel: +32-3-265.10.02 (Secretary VIB8: Mrs. Gisèle Smeyers)
Fax: +32-3-265.10.12
E-mail: mailto:Thomas.Geuens@molgen.vib-ua.be
http://www.molgen.vib-ua.be/









From: technical@abcam.com [mailto:technical@abcam.com]
Sent: Tuesday, November 27, 2012 5:20 PM
To: Geuens Thomas
Subject: Reply from Abcam to your enquiry regarding ab37969 [CCE4396586]



















https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header









Dear Thomas

We have an answer to your inquiry:

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.

Although the other antibody has worked well using this procedure, individual antibodies will often require optimization. I would like to offer some suggestions to help optimize the results from ab37969. I would also appreciate if you can confirm some details of the procedure.

1. I can recommend to consider trying reducing and denaturing conditions. This means the antibody will be in the correct conformation for detection by the antibody and at the correct molecular weight. We recommend to use reducing and denaturing conditions in WB unless it recommends non denaturing or native conditions on the datasheet.

2. Could you confirm if the transfer of protein ot the membrane been assessed using a loading control on the blots on which ab37969 were used?

3. What dilutions of antibody have been tried? I can recommend to increase the concentration in order to help increase the signal.

4. Is the current vial of secondary antibody working well with other primary antibodies?

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.




Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=4396586


Best regards,
Kate

Kate Hayes
Scientific Support Supervisor
Abcam plc
https://www.abcam.com


Your original inquiry to Abcam:


















Thanks for replying. See attached picture and filled in form at end of this email.



Thanks a lot.



Best and friendly greetings,



Thomas Geuens



----------------------------------------------------------------------
Thomas Geuens, PhD Student
VIB - Department of Molecular Genetics
Peripheral Neuropathy Group
University of Antwerp - CDE
Parking P4, Building V, Room V1.17
Universiteitsplein 1
BE-2610 Antwerpen
Belgium

Tel: +32-3-265.10.27
Tel: +32-3-265.10.02 (Secretary VIB8: Mrs. Gisèle Smeyers)
Fax: +32-3-265.10.12
E-mail: mailto:Thomas.Geuens@molgen.vib-ua.be
http://www.molgen.vib-ua.be/









From: mailto:technical@abcam.com [mailto:technical@abcam.com]
Sent: Tuesday, November 27, 2012 12:16 PM
To: Geuens Thomas
Subject: Reply from Abcam to your enquiry regarding ab37969 [CCE4394871]



















https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header









Dear Thomas

We have an answer to your inquiry:

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 for WB and for mouse and human 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.

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

Order Details
Antibody code:
ab37969
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Very weak to no signal
Lot number
GR53724-1
Purchase order number
or preferably Abcam order number:
70102629, ordered by Eric Patteet around the date of 5 November 2012

General Information
Antibody storage conditions (temperature/reconstitution etc)
-20°C

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

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Human SH-SY5Y and mouse Spinal Cord

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
RIPA – roche protease inhibitor tablet

Amount of protein loaded
Human SH-SY5Y (20µg) mouse Spinal Cord (50µg)

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Non-reducing, 4-10% NuPage gel

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
90min transfer time, 5% milk diluted in PBS+0.01% Tween-20

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
ab37969 (1/500 in 5% Milk diluted in PBS+0.01% Tween-20)

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Rabbit-HRP (1/500 in 5% Milk diluted in PBS+0.01% Tween-20)

Detection method (ECL, ECLPlus etc.)
ECL Plus

Positive and negative controls used (please specify)
Positive control = anti-eEF1A2 antibody from Abnova (see attached picture)


Optimization attempts (problem solving)
How many times have you tried the Western?
4 times trying different dilutions of primary and secondary antibodies. Also I tried different species and amount of proteins loaded on gel (see attached picture)


Have you run a "No Primary" control?
Yes (see attached picture)


Do you obtain the same results every time?
yes


e.g. are the background bands always in the same place?
Yes, if there is background

What steps have you altered?
Antibody dilutions, amount of protein loaded on gel, tried lysates from different species

Additional Notes:
(see attached picture)

Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.



Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=4394871


Best regards,
Kate

Kate Hayes
Scientific Support Supervisor
Abcam plc
https://www.abcam.com


Your original inquiry to Abcam:










Dear Sir/Madam,



Recently I bought an anti human eEF1A1 (n° 37969) from your company. So far I’ve been trying to optimize the antibody in order to get a proper signal. Until now this was not successful, unfortunately. I’ve tried the antibody on the human SH-SY5Y cell line and mouse spinal cord where I loaded 10 and 50µg of proteins respectively. For the mouse tissue I also took along an anti human eEF1A2 from Abnova as a positive control. As you can see on the image attached to this mail the positive control works and gives a nice band around 50 kDa but the Abcam antibody fails. On the right side of the blot you can see that the Abcam antibody also fails with cell extracts. According to the Abcam datasheet 10µg of protein is enough to detect eEF1A1 in both used lysates, using a 1/1000 dilution of the antibody (https://www.abcam.com/eef1a1-eef1a2-eef1al3-antibody-ab37969.html ). The dilution I used was 1/500 for primary as well as for secondary antibodies.



I already bought a lot Abcam antibodies without any problems and with great satisfaction! I was wondering if you can help me with this issue and I hope we can find out a solution.



Best regards and friendly greetings.



Thomas Geuens



----------------------------------------------------------------------
Thomas Geuens, PhD Student
VIB - Department of Molecular Genetics
Peripheral Neuropathy Group
University of Antwerp - CDE
Parking P4, Building V, Room V1.17
Universiteitsplein 1
BE-2610 Antwerpen
Belgium

Tel: +32-3-265.10.27
Tel: +32-3-265.10.02 (Secretary VIB8: Mrs. Gisèle Smeyers)
Fax: +32-3-265.10.12
E-mail: mailto:Thomas.Geuens@molgen.vib-ua.be
http://www.molgen.vib-ua.be/








Abcam Customer Services and Scientific Support Team
https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body

[CCE4394871]














Discover more at abcam.com




























Abcam Customer Services and Scientific Support Team
https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body

[CCE4396586]













Discover more at abcam.com

Read More
Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

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.

Although the other antibody has worked well using this procedure, individual antibodies will often require optimization. I would like to offer some suggestions to help optimize the results from ab37969. I would also appreciate if you can confirm some details of the procedure.

1. I can recommend to consider trying reducing and denaturing conditions. This means the antibody will be in the correct conformation for detection by the antibody and at the correct molecular weight. We recommend to use reducing and denaturing conditions in WB unless it recommends non denaturing or native conditions on the datasheet.

2. Could you confirm if the transfer of protein ot the membrane been assessed using a loading control on the blots on which ab37969 were used?

3. What dilutions of antibody have been tried? I can recommend to increase the concentration in order to help increase the signal.

4. Is the current vial of secondary antibody working well with other primary antibodies?

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 for WB and for mouse and human 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.

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

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:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Answer

Thank you for your reply, and I apologize again for this error. I found your order *** from October 2008, and I have asked our finance department to issue credit note #***. 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. To 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. 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. The cross-reactivity with eEF1A2 and eEF1A3 was predicted based on sequence homology and then confirmed by Western blot against the recombinant proteins. I hope this information is helpful in interpreting your results, but please let me know if you need any further information and I'll be happy to help.

Read More

Answer

Thank you for your reply. The PO# on this order is 1244827. I appreciate you forwarding the email to the end user. Please let me know if you need any more information from me.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549 lung cells)
Specification
A549 lung cells
Fixative
Formaldehyde
Permeabilization
Yes - 1% TX-100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. David Matthews

Verified customer

Submitted Apr 07 2009

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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