Overview

  • Product name
  • Description
    Goat polyclonal to CCR5
  • Host species
    Goat
  • Specificity
    This antibody binds to CCR 5 receptors on human peripheral blood leukocytes as determined by immunocytochemistry.
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, ICCmore details
  • Species reactivity
    Reacts with: Human, Macaque monkey
  • Immunogen

    Synthetic peptide:

    YQVSSPIYDINYYTSEPCQKINVKQIAA

    , corresponding to N terminal amino acids 3-30 of Human CCR5.

  • Positive control
    • Human peripheral blood cells or paraffin sections of spleen.

Properties

Applications

Our Abpromise guarantee covers the use of ab1673 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
IHC-Fr Use at an assay dependent dilution.
WB 1/1000.
IHC-P 1/300. with biotinylated secondary and streptavidin-HRP.
ICC 1/300.

Target

  • Function
    Receptor for a number of inflammatory CC-chemokines including MIP-1-alpha, MIP-1-beta and RANTES and subsequently transduces a signal by increasing the intracellular calcium ion level. May play a role in the control of granulocytic lineage proliferation or differentiation. Acts as a coreceptor (CD4 being the primary receptor) for HIV-1 R5 isolates.
  • Tissue specificity
    Highly expressed in spleen, thymus, in the myeloid cell line THP-1, in the promyeloblastic cell line KG-1A and on CD4+ and CD8+ T-cells. Medium levels in peripheral blood leukocytes and in small intestine. Low levels in ovary and lung.
  • Involvement in disease
    Genetic variation in CCR5 is associated with suseptibility to diabetes mellitus insulin-dependent type 22 (IDDM22) [MIM:612522]. A multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These derangements result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Post-translational
    modifications
    Sulfated on at least 2 of the N-terminal tyrosines. Sulfation contributes to the efficiency of HIV-1 entry and is required for efficient binding of the chemokines, CCL3 and CCL4.
    O-glycosylated, but not N-glycosylated. Ser-6 appears to be the major site. Also sialylated glycans present which contribute to chemokine binding. Thr-16 and Ser-17 may also be glycosylated and, if so, with small moieties such as a T-antigen.
    Palmitoylation in the C-terminal is important for cell surface expression, and to a lesser extent, for HIV entry.
    Phosphorylation on serine residues in the C-terminal is stimulated by binding CC chemokines especially by APO-RANTES.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • AM4 7 antibody
    • C C chemokine receptor type 5 antibody
    • C C CKR 5 antibody
    • C-C chemokine receptor type 5 antibody
    • C-C CKR-5 antibody
    • C-C motif chemokine receptor 5 A159A antibody
    • CC Chemokine Receptor 5 antibody
    • CC Chemokine Receptor Type 5 antibody
    • CC CKR 5 antibody
    • CC-CKR-5 antibody
    • CCCKR 5 antibody
    • CCCKR5 antibody
    • CCR 5 antibody
    • CCR-5 antibody
    • CCR5 antibody
    • CCR5 chemokine (C C motif) receptor 5 antibody
    • CCR5_HUMAN antibody
    • CD 195 antibody
    • CD195 antibody
    • CD195 Antigen antibody
    • Chemokine C C motif receptor 5 antibody
    • Chemokine receptor CCR5 antibody
    • CHEMR13 antibody
    • CKR 5 antibody
    • CKR5 antibody
    • CMKBR 5 antibody
    • CMKBR5 antibody
    • FLJ78003 antibody
    • HIV 1 Fusion Coreceptor antibody
    • HIV-1 fusion coreceptor antibody
    • HIV1 fusion coreceptor antibody
    • IDDM22 antibody
    • MIP-1 alpha receptor antibody
    see all

Images

  • Immunohistochemistry using ab1673 on a section of normal human spleen 

References

This product has been referenced in:
  • Subramanian S  et al. TREM-1 associated macrophage polarization plays a significant role in inducing insulin resistance in obese population. J Transl Med 15:85 (2017). IHC-P ; Human . Read more (PubMed: 28454543) »
  • El-Hage N  et al. HIV-1 coinfection and morphine coexposure severely dysregulate hepatitis C virus-induced hepatic proinflammatory cytokine release and free radical production: increased pathogenesis coincides with uncoordinated host defenses. J Virol 85:11601-14 (2011). IF . Read more (PubMed: 21900165) »
See all 6 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for contacting us.
I am sorry that the vial you received did not contain the full amount of the product. We are very careful to measure out exactly the stated quantity, however, sometimes during delivery the cap can become loosened and some loss of product can occur. This is very rare but I sincerely apologize for the inconvenience this has caused you. I have issued a free of charge replacement vial with the order number of 1058239 (PO number FOCR MDS614616).
Please do not hesitate to contact us if you need anything further.

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Answer

Thank you for your enquiry. I can confirm that ab1673 recognizes a band of approximately 40kDa. and that CCR5 has a predicted molecular weight of 40524Da. If you are continually obtaining a band of 37KDa may I suggest that you fill in our technical questionnaire. This will enable our technical team to better deal with your technical query and facilitate a quick resolution to your problem. I have added the technical questionnaire link for this antibody below. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=1673&mode=questionaire I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

The image supplied shows a paraffin section of normal human spleen tissue. I suspect that the blocking step with the Sniper reagent may be a problem. Here is the protocol that we recommend. Immunohistochemistry on Paraffin sections: Paraffin sections of tissues fixed in 10% formalin are used. Preparation of slides: Place slides in 60 C oven overnight after cutting. Then follow the procedure below: 1) xylene 2x10min 2) 100% alcohol 2x10min 3) 95% alcohol 5 min 4) 80% alcohol 5 min 5) rinse in distilled water 2x2 min 6) antigen retrieval if necessary 7) Immunohistochemistry Antigen Retrieval: Sections were treated with antigen retrieval solution and steamed for 20 minutes followed by cooling for 20 minutes. Immunohistochemistry: Block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. Wash the section with PBS and incubate with primary antibody (anti-chemokine receptor) at recommended dilution for 60 minutes at RT. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L at 1-200 dilution for 30 minutes. Wash the slides and incubate with HRP-Streptavidin 1:400 dilution for 30 minutes. Wash the slides and incubate with DAB for 5-7 minutes at RT. Wash slides with PBS and counter stain with hematoxylin. I hope that this information is helpful. Please contact me if you have any futher questions or concerns.

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Answer

Thank you for your email. I'm not familiar with the universal immunoenzyme polymer method and so I cannot comment as to how it compares with the ABC method. As you are using the Vectastain Universal Quick Kit, I do suggest following their suggestions. Please contact us again if you have any additional questions.

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Question

Thank you for your reply.We looked your home page again after we got your reply,but we didn't understand much abut it.So we decided to answer your form of questions. We would like to you look them and return your opinion. >. Can you please specify which CCR5 antibody you >are using? Please indicate the Abcam code number (ab....). ...ab1673 >Also, I have some general questions, the answers to which will enable me to >investigate this matter as quickly as possible: 1. Please describe the problem (high background, no staining etc). ...It has no staining about lymph cells but several granulocytes were stained. 2. On what material are you testing the antibody in IHC? •Species?... human •Cell line? •Tissue? ....Paraffin embeded sections. 3. How did you fix the samples? •Ethanol, methanol •Acetone •Paraformaldehyde •Other ...We used formaldehyde. 4. Did you apply antigen retrieval step? •Enzymatic method •Heat mediated technique...... we used EDTA buffer(pH8.0) 40 minutes (95? microwave). •Other 5. How did you block the unspecific binding sites? ...We used 3%H2O2 methyl alcohol. 6. Primary antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? ...We diluted it by PBS buffer. •Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)?...We incubeted it over night, after that we rinsed three times by PBS buffer. 7. Secondary antibody •What secondary antibody are you using? ...We used Histofine Simple Stain MAX-PO(MULTI). •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody?...not deluted •Incubation, wash steps?...30 minutes •Do you know whether the problems you are experiencing come from the secondary?...No, we don't. •What detection method are you using? 8. Background staining •Please provide an image of your staining 9. Which detection system did you use? 10. Did you apply positive and negative controls along with the samples?Please specify. ...lymph node and palate tonsil. All cells of both section were no stained. 11. Optimization attempts •How many times have you tried the IHC?...twice •Do you obtain the same results every time?...The first time, all cells were negative. The secound time, several granulocytes were stained. •What steps have you altered?...The first time, we used heat-induced epitope retrieval in citrate buffer for 25minutes. The secound time, we used EDTA buffer(pH8.0). We used this method deparaffinized,rehydrated, ? ?mM EDTA (pH 8.0) with high temperature( 95? microwave). ? rins PBS buffer ? 3% H2O2 methanol for 15 minutes ? rins 3 times with PBS buffer 5 minutes ? Primary antibody( ab1673) ( diluted it by PBS buffer(×300) ) over-night at room temperature ? rins 3 times with PBS buffer 5 minutes ? secondary antibody dirrect in Histofine Simple Stain MAX-PO(MULTI) kit 30minutes at room temperature ? rins 3 times with PBS buffer 5 minutes ? DAB (DAKO) within 10 minutes ? rins distilled water ? hematoxylin stain (for nucleus)

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Answer

Thank you for the details that you have provided. I have a few more questions for you that will help me to troubleshoot this problem: 1. I was unable to open the attachments that you sent along with your email. You mentioned in your email that "All cells of both section were no stained." Does that mean that you are not seeing any staining at all with this antibody? 2. What dilutions of ab1673 have you tried? It is recommended to use ab1673 at a dilution of 1:300 in IHC-P. 3. Have you done a positive control? Ab1673 was tested and characterized for IHC using paraffin sections of human spleen, which is recommended as a positive control. 4. What was the batch number that you received (it is located on the vial)? Also, did you order directly from Abcam or through one of our distributors? Thank you again, and I look forward to hearing from you.

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