Overview

  • Product name

  • Description

    Rabbit polyclonal to Iba1
  • Host species

    Rabbit
  • Specificity

    In WB, we recommend blocking in milk. Blocking with BSA gives high background. If looking for a monoclonal anti-Iba1 alternative we can recommend our RabMAb ab178846
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cat, Human
  • Immunogen

    Synthetic peptide from the C terminal amino acids of Human Iba1 (NM_001623.3).

  • Positive control

    • Iba1 (Human) full length recombinant protein; Rat brain lysate; Human fetal lymphocytes; spleen tissue lysate; THP1, C6, NR8383 whole cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab108539 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 1/500 - 1/1000. Predicted molecular weight: 17 kDa.

We recommend blocking in 3-5% milk. Blocking with BSA gives high background.

IHC-P 1/100 - 1/500.

Target

  • Function

    Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.
  • Tissue specificity

    Detected in T-lymphocytes and peripheral blood mononuclear cells.
  • Sequence similarities

    Contains 2 EF-hand domains.
  • Post-translational
    modifications

    Phosphorylated on serine residues.
  • Cellular localization

    Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
  • Information by UniProt
  • Database links

  • Alternative names

    • AIF 1 antibody
    • AIF-1 antibody
    • Aif1 antibody
    • AIF1 protein antibody
    • AIF1_HUMAN antibody
    • Allograft inflammatory factor 1 antibody
    • Allograft inflammatory factor 1 splice variant G antibody
    • allograft inflammatory factor-1 splice variant Hara-1 antibody
    • balloon angioplasty responsive transcription antibody
    • BART 1 antibody
    • G1 antibody
    • G1 putative splice variant of allograft inflamatory factor 1 antibody
    • IBA 1 antibody
    • IBA1 antibody
    • interferon gamma responsive transcript antibody
    • Interferon responsive transcript 1 antibody
    • interferon responsive transcript factor 1 antibody
    • Ionized calcium binding adapter molecule 1 antibody
    • Ionized calcium-binding adapter molecule 1 antibody
    • ionized calcium-binding adapter molecule antibody
    • IRT 1 antibody
    • IRT1 antibody
    • Microglia response factor antibody
    • MRF1 antibody
    • Protein g1 antibody
    see all

Images

  • IHC image of Iba1 staining in normal human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab108539, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of Iba1 staining in normal mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab108539, 1/1000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of Iba1 staining in normal rat brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab108539, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

     

  • All lanes : Anti-Iba1 antibody (ab108539) at 1/1000 dilution

    Lane 1 : Human Iba1 recombinant protein at 0.1 µg
    Lane 2 : HEK293 whole cell lysate at 20 µg
    Lane 3 : A431 whole cell lysate at 20 µg
    Lane 4 : NIH3T3 whole cell lysate at 30 µg
    Lane 5 : Human spleen tissue lysate at 20 µg
    Lane 6 : Mouse spleen tissue lysate at 30 µg
    Lane 7 : Rat spleen tissue lysate at 30 µg
    Lane 8 : U937 whole cell lysate at 30 µg
    Lane 9 : MOLT4 whole cell lysate at 20 µg
    Lane 10 : THP1 whole cell lysate at 30 µg
    Lane 11 : THP1 whole cell lysate, PMA treated at 30 µg
    Lane 12 : Raw 264.7 whole cell lysate at 30 µg
    Lane 13 : C6 whole cell lysate at 30 µg
    Lane 14 : NR8383 whole cell lysate at 30 µg
    Lane 15 : NIH3T3 whole cell lysate - BLOCKED IN 5% BSA at 30 µg
    Lane 16 : Human spleen tissue lysate - BLOCKED IN 5% BSA at 20 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17 kDa


    Exposure time: 3 minutes


    Lanes 1-14: Blocked in 3% milk for 1 hour (RT). Lanes 15-16: Blocked in 5% BSA for 1 hour (RT).

    Abcam recommends blocking in milk for cleaner blots with reduced background, in comparison to BSA. 

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab108539 (anti-Iba1 antibody; 1/1000) for 18 hours at 4°C. Antibody binding was detected using HRP-labelled anti-Rabbit IgG for 1 hour at room temperature and visualised using ECL development solution ab133406.

  • Anti-Iba1 antibody (ab108539) at 1/500 dilution + Rat brain lysate

    Predicted band size: 17 kDa

  • ab108539, at 1/100 dilution, staining Iba1 in formalin-fixed, paraffin-embedded Human fetal lymphocytes by Immunohistochemistry.

References

This product has been referenced in:

  • Tyrtyshnaia A  et al. Neuroinflammation and adult hippocampal neurogenesis in neuropathic pain and alkyl glycerol ethers treatment in aged mice. Int J Mol Med 43:2153-2163 (2019). Read more (PubMed: 30896810) »
  • Aricioglu F  et al. Antidepressant-like Effects Induced by Chronic Blockade of the Purinergic 2X7 Receptor through Inhibition of Non-like Receptor Protein 1 Inflammasome in Chronic Unpredictable Mild Stress Model of Depression in Rats. Clin Psychopharmacol Neurosci 17:261-272 (2019). Read more (PubMed: 30905126) »
See all 18 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for contacting Abcam.

So that I can better help you with this, would you be able to provide me with a copy of the image that you are getting and also the protocol that you are using, in particular could you include the following information:

1 - What sample type are you using?

2 - How much sample are you loading?

3- What type of blocking agent are you using?

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

5 - How long do you boil your samples for?

I look forward to your reply and helping you resolve this issue.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: MW pH9
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1500µg/mL
Fixative
Paraformaldehyde

Gaia Brezzo

Verified customer

Submitted Dec 16 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Dec 01 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 30 2015

Answer

Thank you for contacting us.

I can confirm that antibody ab108539 could be used for detection of Iba1 in microglia cells of mouse brain. This assumption is however based on the expression of this protein in mouse microglia cells; we unfortunately haven't tried this in our lab however it would work.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for taking the time to contact us. I am sorry to hearthe customer hashad difficulty obtaining satisfactory results from these threeantibodies.

I would like to reassure you that 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 could also provide images which would help us to assess the results.

Please note Ihave sent the questionnaire for IHC and ICC. If the customer has used another application, please let me know and I can send the correct form so we can obtain the relevant information required.

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



Order Details
Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


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


Positive and negative controls used (please specify)


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



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

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which woudl help us to assess the results

Read More

Answer

Here is a quite detailed protocol for performing paraffin embedded immunohistochemical staining, which has the buffer composition for Tris-EDTA pH 9 which I suggested trying: https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf Tris-EDTA buffer: 10 mM Tris Base, 1 mM EDTA, 0.05% Tween 20, pH9.0 Its quite a useful guide, especially when troubleshooting for a new experiment. I hope it is of help to you. Have a nice weekend.

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Answer

Thank you for contacting us again. As discussed over the phone, I have arranged for the free of charge replacement with ab5076 to be delivered to you at the following address (taken from your previous order): The new order number is: This should be with you tomorrow. I hope this one works better for you. Although your collegues have said that they have not required any antigen retrieval, several of our customers have used citrate mediated antigen retrieval so you may need to optimise when using your particular samples. I was also wrong to say that this antibody has not yet been tested in mouse samples, several of our customers have used it for IHC with mouse tissue and I will amend this information on the datasheet. As discussed I have also attached a questionnaire which it would be very helpful if you could fill out. This will allow me to investigate this case internally and initiate any additional testing where necessary. I look forward to receiving your reply.

Read More

Answer

Thank you for your enquiry. 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, any Iba1 antibody tested in the species and application you are using should be suitable for your applications. I can confirm that both ab5076 and ab108539 have been tested and guaranteed for IHC-P. 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. If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet). Regarding the secondary antibody, ab6884 Donkey polyclonal Secondary Antibody to Goat IgG (Biotin) is tested and guaranteed in IHC-P. As an anti-goat antibody, it would be suitable for use with Anti-Iba1 antibody (ab5076), as this primary antibody is raised in goat. With regards to ab108539 Anti-Iba1 antibody, this is a rabbit polyclonal, and therefore requires an anti-rabbit secondary antibody and ab6884 would not be a suitable secondary to use with ab108539.. I hope this information will be helpful to you. Should you have any further questions regarding your search and selection of products, please do not hesitate to ask.

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