Overview

  • Product name
  • Description
    Goat polyclonal to KIF4A
  • Host species
    Goat
  • Tested applications
    Suitable for: IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Pig
  • Immunogen

    Synthetic peptide:

    SFFSGCSPIEEEAH

    , corresponding to C terminal amino acids 1219-1232 of Human KIF4A.

Properties

Applications

Our Abpromise guarantee covers the use of ab3815 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
IP Use at an assay dependent concentration.
WB 1/100. Can be blocked with Human KIF4A peptide (ab22955).
ICC/IF 1/100.

Target

  • Function
    Motor protein that translocates PRC1 to the plus ends of interdigitating spindle microtubules during the metaphase to anaphase transition, an essential step for the formation of an organized central spindle midzone and midbody and for successful cytokinesis. May play a role in mitotic chromosomal positioning and bipolar spindle stabilization.
  • Tissue specificity
    Highly expressed in hematopoietic tissues, fetal liver, spleen, thymus and adult thymus and bone marrow. Lower levels are found in heart, testis, kidney, colon and lung.
  • Sequence similarities
    Belongs to the kinesin-like protein family. Chromokinesin subfamily.
    Contains 1 kinesin-motor domain.
  • Cellular localization
    Nucleus matrix. Cytoplasm > cytoskeleton > spindle. Midbody. Chromosome. Not present in the nucleolus. In early mitosis, associated with the mitotic spindle, in anaphase, localized to the spindle midzone and, in telophase and cytokinesis, to the midbody. In late cytokinesis, found in the center of the midbody. Associated with chromosomes at all stages of mitosis.
  • Information by UniProt
  • Database links
  • Alternative names
    • Chromokinesin antibody
    • Chromokinesin-A antibody
    • Chromosome associated kinesin KIF4A antibody
    • Chromosome-associated kinesin KIF4A antibody
    • FLJ12530 antibody
    • FLJ12655 antibody
    • FLJ14204 antibody
    • FLJ20631 antibody
    • HSA271784 antibody
    • KIF4 antibody
    • KIF4 G1 antibody
    • KIF4A antibody
    • KIF4A_HUMAN antibody
    • KIF4G1 antibody
    • Kinesin family member 4A antibody
    see all

Images

  • ab3815 (1/100) stainning KIF4A in HeLa Cells (blue). Cells were fixed with Methanol and BSA was used as blocking agent. Please refer to abreview for further experimental details.

    See Abreview

  • All lanes : Anti-KIF4A antibody (ab3815) at 1/100 dilution

    Lane 1 : Whole cell lysate prepared from HeLa cells synchronized with Nocodazole for 12 hours.
    Lane 2 : As Lane one but with KIF4A siRNA treatment as negative control.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP conjugated anti goat at 1/5000 dilution

    Developed using the ECL technique.

    Observed band size: 140 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes

References

This product has been referenced in:
See all 5 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

Thank you for taking the time tocontact us. I am sorry to hearthe customer has had difficulty obtaining satisfactory results from this second vial of antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time the customer has spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there would be no further tips to provide on this occasion to help improve the results andI apologize for the inconvenience. I'm pleased to offer you acredit note or alternative replacementin compensation, which may be more appropriate in this case than another vial of the same antibody.

In addition, I would appreciate if you could confirm the following information which will help with our quality monitoring:

1. Please confirm if the same type of samples have been used in this case (HeLa cells).

2. Has the same procedure (with the suggestions previously provided) been used?

3. What is the problem with the results? Is there high background in WB and ICC again?

4. Has the customer tried fresh samples?

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab3815 has been added to your order number [BO] 88841. This has Abcam order reference 1112739.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

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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 suggestthe customer has 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 the customers concerns and if they prefer a credit note in this case, I will be pleased to arrange this for them.

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

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Question
Answer

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

I am sorry to hear the suggestions made have not improved the results on this occasion. 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.

I appreciate the time you have spent on these experiments, and would like to offer a refund or credit note in compensation (providing the product has been purchased in the last 6 months). In order to arrange this, I would appreciate if you could confirm your order number and date of purchase as previously requested?

Thank you for your continued cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question

Product code: 3815
Lot number: GR86868-1

Inquiry: Antibody code: ab3815
Batch number: GR86868-1
General Information:
Antibody storage conditions (temperature/reconstitution etc) -20deg
Description of the problem (high background, low signal, non-specific staining etc.) non-specific staining, no relevant signal
Sample (Species/Tissue/Cell Type/Cell Line etc.) HELA cells
Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
Tried both Methanol/Acetone/ or Paraformaldehyde
Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
Permeabilization step Aceton for Methanol, Triton for formaldehyde
Blocking conditions (Buffer/time period, Blocking agent etc.) 5% BSA, 1h
Primary Antibody (Diluent/Dilution/Incubation time, Wash step) 1h RT 1:100, 3 washes PBST
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
Wash step) Alexa anti goat 594, 1:200, 1h RT
Detection method Observer. Z1 Microscope (Zeiss)
Positive and negative controls used (please specify) negative control – no primary Optimization attempts (problem solving):
How many times have you tried the IHC? 2 times
Have you run a "No Primary" control? yes Do you obtain the same results every time? yes
What steps have you altered?
Additional Notes
Document Attachment: images of the results may be very helpful. If you wish to add it to the complaint form, please attach them to the reply e-mail.

Antibody code: ab3815 Batch number: GR86868-1
Antibody storage conditions (temperature/reconstitution etc) -20
Description of the problem (high background, wrong band size, more bands, no band etc.) no relevant band, many non-specific bands
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) HELA cell extract, 293-T cell extract, HELA and 293-T synchronized with Nocodazole for 12h lysates
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) RIPA buffer + protease inhibitors (merk)
Amount of protein loaded 30microg
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) reducing gel 10%
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) 1h 20min semi-dry transfer, TGB +20% MetOH transfer buffer, blocking with 5% milk
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) 1:100 in 1% BSA solution, 1h RT or ON 4deg, 6 washes with PBST
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Anti goat 1:5000 (Jackson), 1% BSA, 1h RT, 6 washes with PBST
Detection method (ECL, ECLPlus etc.) ECL Positive and negative controls used (please specify) OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
How many times have you tried the Western? 3 times Have you run a "No Primary" control?
Do you obtain the same results every time? e.g. are the background bands always in the same place? yes
What steps have you altered? ON incubation with ab, cell syncronization Additional Notes

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Answer

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

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.

I would like to reassure you that ab3815 is tested and covered by our 6 month guarantee for use in WB, ICC-IF and inhuman samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if youare able to confirm some further details:

ICC-IF

1. Has the quality of the sample been assessed, for example what was the% confluency before staining, and was a spare sample checked for viability using trypan blue count before staining?

2. Fixation: To prevent over fixation, whichever method is used, I can suggest this should be for 10 minutes only if this has not already been tried. PFA should be 4% only.

3. Please provide further information regarding the permeabilization. I would suggest to try 0.2% triton, 10 minutes only.

4. I can recommend to consider reducing the concentration to 1:500 to reduce the background signal.

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

WB

1. I can suggest to try reducingthe concentration to 1:500 to reduce the background signal. Incubate overnight 4oC.

2. Could you confirm if the quality of sample been assessed using a loading control?

3. Have the samples reduced and denatured?

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

General:

1. Please confirm the order number and date of purchase.

2. 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.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cell line)
Specification
HeLa cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - triton .15
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 09 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa cells)
Total protein in input
10 µg
Specification
HeLa cells
Treatment
500nM nocodazole for 12 hrs
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted May 28 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
10 µg
Specification
HeLa
Treatment
Nocodazole synchronized for 12hrs
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Mar 23 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Dec 03 2008

Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. We do experience occasional batch variation. This often means that it may be necessary for our customers to perform an element of optimisation in their experiments when receiving the antibody. On this occasion I would like to recommend that your customer increases the mass of protein that they are loading on the gel as we typically recommend that 20-30ug of protein is loaded. I would also recommend that the antibody is applied at a dilution of 1:100; as detailed on our datasheet using an overnight incubation. Taken in combination this is very likely to improve the results that your customer has been obtaining significantly. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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

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