Overview

  • Product name
    Anti-CD64 antibody [10.1]
    See all CD64 primary antibodies
  • Description
    Mouse monoclonal [10.1] to CD64
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IP, IHC-P, IHC-Fr, Functional Studies, ICC, Flow Cyt, Blockingmore details
  • Species reactivity
    Reacts with: Human, Non human primates
  • Immunogen

    Rheumatoid synovial fluid cells and fibronectin purified Human monocytes.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    ab119843 is purified from cell culture supernatant by protein-A affinity chromatography. Purity is > 95% by SDS-PAGE.
  • Clonality
    Monoclonal
  • Clone number
    10.1
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab119843 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 at an assay dependent concentration. Predicted molecular weight: 72 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Functional Studies Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
Flow Cyt Use a concentration of 5 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Blocking Use at an assay dependent concentration.

Target

  • Function
    High affinity receptor for the Fc region of immunoglobulins gamma. Functions in both innate and adaptive immune responses.
  • Tissue specificity
    Monocyte/macrophage specific.
  • Sequence similarities
    Belongs to the immunoglobulin superfamily. FCGR1 family.
    Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
  • Post-translational
    modifications
    Phosphorylated on serine residues.
  • Cellular localization
    Cell membrane. Stabilized at the cell membrane through interaction with FCER1G.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD 64 antibody
    • CD64 antibody
    • CD64 antigen antibody
    • CD64A antibody
    • CD64b antibody
    • CD64c antibody
    • Fc fragment of IgG high affinity Ia receptor antibody
    • Fc fragment of IgG high affinity Ib receptor antibody
    • Fc fragment of IgG high affinity Ic receptor antibody
    • Fc fragment of IgG, high affinity Ia, receptor (CD64) antibody
    • Fc fragment of IgG, high affinity Ia, receptor for (CD64) antibody
    • Fc fragment of IgG, high affinity Ia, receptor for antibody
    • Fc fragment of IgG, high affinity Ib, receptor (CD64) antibody
    • Fc fragment of IgG, high affinity Ib, receptor for antibody
    • Fc fragment of IgG, high affinity Ic, receptor (CD64) antibody
    • Fc fragment of IgG, high affinity Ic, receptor (CD64), pseudogene antibody
    • Fc fragment of IgG, high affinity Ic, receptor for antibody
    • Fc gamma receptor antibody
    • Fc gamma receptor I A1 antibody
    • Fc gamma receptor I antibody
    • Fc gamma receptor I B2 antibody
    • Fc gamma RI antibody
    • Fc gamma RIA antibody
    • Fc gamma RIB antibody
    • Fc of IgG high affinity Ia receptor antibody
    • Fc-gamma RI antibody
    • Fc-gamma RIA antibody
    • Fc-gamma RIC antibody
    • FCG 1 antibody
    • FCG1 antibody
    • FcgammaRI antibody
    • FcgammaRIa antibody
    • FCGR 1 antibody
    • FCGR1 antibody
    • FCGR1_HUMAN antibody
    • FCGR1A antibody
    • FCGR1B antibody
    • FCGR1C antibody
    • FcRI antibody
    • FcRIB antibody
    • FCRIC antibody
    • FLJ18345 antibody
    • HFcgammaRIB antibody
    • High affinity immunoglobulin gamma Fc receptor I antibody
    • High affinity immunoglobulin gamma Fc receptor IB antibody
    • IGFR 1 antibody
    • IGFR1 antibody
    • IGFRB antibody
    • IGFRC antibody
    • IgG Fc receptor I antibody
    • IgG Fc receptor IB antibody
    • IgG Fc receptor IC antibody
    • Immunoglobulin G Fc receptor I antibody
    • Immunoglobulin G Fc receptor IB antibody
    • Immunoglobulin G Fc receptor IC antibody
    • MGC137713 antibody
    see all

Images

  • Surface staining of CD64 in human peripheral blood with anti-CD64 (10.1) purified , GAM-APC.
  • Overlay histogram showing THP1 cells stained with ab119843 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab119843, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 80% methanol (5 min) fixed THP1 cells used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Zhang L  et al. C-reactive protein exacerbates epithelial-mesenchymal transition through Wnt/ß-catenin and ERK signaling in streptozocin-induced diabetic nephropathy. FASEB J 33:6551-6563 (2019). Read more (PubMed: 30794428) »
  • Bajpai G  et al. The human heart contains distinct macrophage subsets with divergent origins and functions. Nat Med 24:1234-1245 (2018). Read more (PubMed: 29892064) »
See all 8 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

I have issued the free of charge antibody ab104273, which should be with you on Monday. The order number is xxxxxx. I hope the suggestions that I have made if not the new antibody will help. I wish you well with your experiments. If you do continue to have problems (or if the staining starts to work) please do let me know.

Read More

Question

Abcam IHC/ICC Questionnaire             It will take approximately 5 minutes to complete the questionnaire.  Please fill-in all fields so that we can better assist you.    1) Abcam product code  ab119843                           2) Abcam order reference number or product batch number   3) Description of the problem – unable to detect CD64 staining in Paraffin embedded lung tissue.  Tried pH6/9 heat pretreatments and two dilutions of the mAb (5&10µg/ml)   4) Sample preparation: Species            Human Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: PFA/formalin fixed paraffin embedded sections Sample preparation   Positive control none Negative control TBS no primary   5) Fixation step N/A Yes/No If yes:  Fixative agent and concentration Fixation time Fixation temperature   6) Antigen retrieval method (if applicable) EDTA pH9, Citrate pH6 (microwave 20 min)   7) Permeabilization method: N/A Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Permeabilizing agent and concentration:     8) Blocking agent (eg BSA, serum…): 3% normal Horse serum Concentration Blocking time 30 min                                         Blocking temperature RT   9) Endogenous peroxidases blocked? (H202(3%) (in MeOH)) 30 min @ RT Endogenous biotins blocked? NO   10) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution 5, 10 µg/ml  Diluent buffer TBS (3% Horse serum)  Incubation time overnight @ 4oC     11)  Secondary antibody:           Species:Horse Reacts against:Mouse Concentration or dilution 1/200 Diluent buffer TBS   Incubation time 30 min Fluorochrome or enzyme conjugate Biotin   12) Washing after primary and secondary antibodies: Buffer – TBSt(0.05%) Number of washes x3 3 min         13) Detection method – vector ABC elite, DAB chromagen   14) How many times have you run this staining? x1 Do you obtain the same results every time? N/A What steps have you altered to try and optimize the use of this antibody? Antigen retrieval and primary mAb dilution   Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.   Images attached    

Read More
Answer

Thank you for providing that extra information, and sorry for the delay in getting back to you. I wanted to make sure that I could help you in the best way possible before responding. I have been in contact with the source of this antibody (ab119843) and unfortunately they have observed that the epitope which this antibody recognises is quite sensitive to paraformaldehyde fixation and results have been mixed. The antigen retrieval method used may therefore have to be optimised for your particular sample. I am sorry for this. Due to the inconvenience already caused to you by the failure of ab23351 I would like to offer you another antibody of ours, free of charge. ab104273 recognises the cytomplasmic region of human FCGR1A (also known as CD64) and has been used previously with Formalin/PFA-fixed paraffin-embedded sections, and as the immunogen used is a synthetic peptide, the epitope may be more amenable to fixation. This way you will be able to optimise with both antibodies and see which one works the best straight away. If you would like me to organise this please let me know. With both antibodies there are a few things I would suggest trying in order to improve the staining currently observed: 1. I would try using enzyme antigen retrieval (such as trypsin or pepsin) in order to unmask the epitope. It would also be worthwhile to try the staining without any antigen retrieval. (If using ab104273 I would also try using the citrate buffer and EDTA buffer you have already been using with ab119843 but with different treatment times, try 5, 10 15 and 20 minutes). 2. I would change they way that you are currently blocking the endogenous peroxidase activity. Some cell surface markers are very sensitive to methanol/hydrogen peroxide quenching which can lead to a reduction in the staining of antigenic site. I would therefore recommend using 0.3 % hydrogen peroxide in PBS. You could also alter when this step is performed. By incubating the primary antibody, then incubating with the hydrogen peroxide this problem can be avoided. Suitable controls will obviously need to be performed to ensure the activity is blocked sufficiently. I hope these suggestions help. If however you find that they do not, please let me know. The antibody you are using is covered by the Abpromise for the application and species you are using, you would therefore be entitled to a replacement antibody or a refund if the results did not improve. Please let me know if you would be interested in receiving ab104273 to try.

Read More

Question
Answer

Thank you for contacting us in regards to the problems experienced with ab119843. As we discussed on the phone, I have attached a questionnaire so that I can gather further information regarding the samples tested and the protocol used. Once I receive the completed questionnaire, I will look at the protocol and see if there are any suggestions I can make that may improve the results. This information will also allow us to investigate this case internally and initiate any additional testing where necessary. I have also requested further information on how the antibody has been used previously with IHC and will share that with you as soon as I have further information. As I mentioned over the phone, the antibody is covered by the Abpromise and if we can't get it to work sufficiently you would be entitled to another replacement or a refund. But hopefully it won't come to that. I look forward to receiving your reply.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up