Overview

  • Product name
  • Description
    Goat polyclonal to CCR7
  • Host species
    Goat
  • Specificity
    Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region. The antibody has been tested in an academic lab on cells transfected with CCR7 receptor, we do not have information regarding its use against endogenous protein.
  • Tested applications
    Suitable for: ELISA, Flow Cyt, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide:

    DPGKPRKNVLVVALLVIFQVC

    , corresponding to amino acids 2-22 of Mouse CCR7.

  • Positive control
    • IHC-P: Mouse spleen, human spleen. WB: cells transfected with CCR7 receptor

Properties

Applications

Our Abpromise guarantee covers the use of ab1657 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
ELISA 1/100000.
Flow Cyt 1/10.

ab37373 - Goat polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P 1/250.
WB 1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).

Target

  • Function
    Receptor for the MIP-3-beta chemokine. Probable mediator of EBV effects on B-lymphocytes or of normal lymphocyte functions.
  • Tissue specificity
    Expressed in various lymphoid tissues and activated B- and T-lymphocytes, strongly up-regulated in B-cells infected with Epstein-Barr virus and T-cells infected with herpesvirus 6 or 7.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • BLR 2 antibody
    • BLR2 antibody
    • C C chemokine receptor type 7 antibody
    • C C CKR 7 antibody
    • C-C chemokine receptor type 7 antibody
    • C-C CKR-7 antibody
    • CC chemokine receptor 7 antibody
    • CC chemokine receptor type 7 antibody
    • CC CKR 7 antibody
    • CC-CKR-7 antibody
    • CCCKR7 antibody
    • CCR 7 antibody
    • CCR-7 antibody
    • Ccr7 antibody
    • CCR7_HUMAN antibody
    • CD 197 antibody
    • CD197 antibody
    • CD197 antigen antibody
    • CDw197 antibody
    • Chemokine (C C motif) receptor 7 antibody
    • Chemokine (C C) receptor 7 antibody
    • Chemokine C C motif receptor 7 antibody
    • Chemokine C C receptor 7 antibody
    • Chemokine receptor 7 antibody
    • Chemokine receptor 7-like protein antibody
    • CMKBR7 antibody
    • EBI 1 antibody
    • EBI1 antibody
    • Ebi1h antibody
    • EBV Induced G Protein Coupled Receptor 1 antibody
    • EBV-induced G-protein coupled receptor 1 antibody
    • Epstein Barr virus induced G protein coupled receptor antibody
    • Epstein Barr virus induced gene 1 antibody
    • Epstein-Barr virus-induced G-protein coupled receptor 1 antibody
    • EVI 1 antibody
    • EVI1 antibody
    • Lymphocyte Specific G Protein Coupled Peptide Receptor antibody
    • MGC108519 antibody
    • MIP 3 beta receptor antibody
    • MIP-3 beta receptor antibody
    • MIP3 Beta Receptor antibody
    see all

Images

  • ab1657, at a dilution of 1/250, staining CCR7 in mouse spleen section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
  • Anti-CCR7 antibody (ab1657) at 1 µg/ml + K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 10 µg

    Secondary
    Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa


    Exposure time: 20 minutes

References

This product has been referenced in:
  • Salim SY  et al. CD83+CCR7- dendritic cells accumulate in the subepithelial dome and internalize translocated Escherichia coli HB101 in the Peyer's patches of ileal Crohn's disease. Am J Pathol 174:82-90 (2009). IHC-P ; Human . Read more (PubMed: 19095953) »
  • Pitkin L  et al. Expression of CC chemokine receptor 7 in tonsillar cancer predicts cervical nodal metastasis, systemic relapse and survival. Br J Cancer 97:670-7 (2007). IHC-P ; Human . Read more (PubMed: 17687340) »
See all 8 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
Sample
Mouse Tissue sections (spleen)
Specification
spleen
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 19 2014

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (spleen, lymph node)
Specification
spleen, lymph node
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C

Mr. Jong Hyung Lim

Verified customer

Submitted Nov 12 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil)
Specification
tonsil
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
methanol/hydrogen peroxide (and I also tried 10% BSA) as blocking agent for 5 minute(s) · Concentration: 2.5%

Abcam user community

Verified customer

Submitted May 30 2007

Question
Answer

Thank you for your patience. I have been in touch with the source of this antibody once again and I have some additional information. Unfortunately at this time I cannot source an alternative lot. However, the originator mentioned the following valuable information. They mentioned that blocking with 2% rabbit serum should be done prior to incubation with the primary antibody. There is no need for dilution of the antibody in 7.5% rabbit serum. Normal rabbit sera contains low levels of auto antibodies to various antigens in their blood. Normal rabbit serum at such high concentration (7.5%) contains a low level of cross-reacting antibodies that may inhibit binding of goat anti-mCCR7. IgG levels in rabbit serum could be 20-30mg/mL. At 7.5% NRS, the IgG level would be 1.5mg/mL in solution combined with approximately 4 ug/mL of goat anti-mCCR7 antibody. I hope this information helps, if this does not improve the results that you are obtaining I would be happy to provide you with a credit note against this purchase.

Read More

Answer

Thank you for taking the time to complete our questionaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody (ab16527). I have checked the procedure used to test this antibody in the laboratory (I have copied this below). Your method follows this very closely, although there are a couple of suggestions I can make to optimise your results. 1. To ensure the sample is not overblocked, I can suggest using just 2% serum for blocking. 2. Can you confirm the constituents of the commercial antigen retreival solution you have used? This antibody has given good results with EDTA buffer heat mediated antigen retrieval for 20 minutes (followed by cooling for 20 minutes). I suggest trying this if you have not done so already. 3. You can increase the signal by increasing the antibody concentration. I suggest trying 1:100. TESTING PROCEDURE: 1.Block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). 2.Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. 3. Wash the section with PBS and incubate with primary antibody at recommended dilution for 60 minutes at RT. 4. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L PUBLISHED PROCEDURE: The following link cites this protocol in the following publication: http://clincancerres.aacrjournals.org/cgi/content/full/11/5/1743 Immunohistochemical Staining. The avidin-biotin complex method was used to detect anti-CCR7, dilution 1:250 Formalin fixed and paraffin-embedded tissues were deparaffinized and subsequently microwaved in EDTA buffer. After preincubation with hydrogen peroxide, avidin/biotin blocking kit, and rabbit serum, the primary antibody was applied for 1 hour at room temperature. After incubation with the secondary antibody (rabbit anti-goat biotinylated), the avidin-biotin complex was added and the enzyme activity visualized with diaminobenzidine. Counterstaining was done with hematoxylin. I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (spleen)
Specification
spleen
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 10%

Dr. Linda Svantesson

Verified customer

Submitted Nov 17 2006

Answer

Thank you for your enquiry. I am sorry to hear that you have been obtaining non-specific staining using this antibody. I have read your technical questionnaire and I have a few comments. Please can you detail the duration of antigen retrieval that you have been performing. We make the recommendation that antigen retrieval is performed for 20 minute as detailed on the antibody datasheet. My concern is that the tissue preparation is being compromised influencing the presentation of the epitope to the antibody thereby causing non-specific staining. I would also appreciate comments with regards whether you have tried no antibody control experiments to control for non-specific staining of this antibody. Secondary antibodies are often discounted as being causal of non-specific staining. I look forward to hearing from you.

Read More

Question
Answer

Thank you for your enquiry. Further to our telephone conversation yesterday afternoon I have been in touch with the source of this antibody. They informed me that by western blotting an actual molecular weight of 43KDa was detected in the following publication. I will e-mail this reference in a follow up e-mail. Walker SR et al. Murine neuroblastoma attenuates dendritic cell cysteine cysteine receptor 7 (CCR7) expression. J Pediatr Surg 40:983-7 (2005). PubMed: 15991182 It may be useful to examine the western blotting approach that was employed in this publication. However, following close examination of this reference it is clear that they do not show a molecular weight standard and the band is cropped in the image, therefore making it difficult to know the ACTUAL migration of the band. Should you continue to have difficulty with this antibody I would encourage you to submit a technical questionnaire by clicking on the link below. This will better enable our technical team to determine the steps that you have taken to optimise this antibody. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=1657&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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Question

I have read your protocol and I don't know which is the difference between yours and mine. Please, Could you help me? Protocol: 1. Please describe the problem (high background, no staining etc). We have high background and no staining in limphocytes. 2. On what material are you testing the antibody in IHC? ? Species? ? Cell line? ? Tissue? Human skin. Paraffin embedded 3. How did you fix the samples? ? Ethanol, methanol ? Acetone ? Paraformaldehyde ? Other Formaldehide 4% (24-48 hours). 4. Did you apply antigen retrieval step? ? Enzymatic method ? Heat mediated technique ? Other -Microwave 2 times, 3 minutes at 400 W. Citrate pH 6. -Steamer 30 minutes EDTA, or TrisEDTA. -Pressures Cooker, citrate pH6, 1 minute. 5. How did you block the unspecific binding sites? BSA 3% 30 minutes 6. Primary antibody ? Specification (in which species was it raised against)? ? At what dilution(s) have you tested this antibody? ? Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? CCR4 Goat. 1:300, 1:100, 1:600. CCR7 Goat. 1:250, 1:500 7. Secondary antibody ? What secondary antibody are you using? ? Specification (in which species was it raised against)? ? At what dilution(s) have you tested this antibody? ? Incubation, wash steps? ? Do you know whether the problems you are experiencing come from the secondary? ? What detection method are you using? Antigoat secondary antibody 1:400. 8. Background staining ? Please provide an image of your staining. 9. Which detection system did you use? SEcondary antibody antigoat (dako) and kwick antibody Shandon. 10. Did you apply positive and negative controls along with the samples? Please specify. None. 11. Optimization attempts ? How many times have you tried the IHC? ? Do you obtain the same results every time? ? What steps have you altered? 5 times. Always the same results.

Read More
Answer

Thank you for your e-mail. We hope these answers are more helpful. 5. How did you block the unspecific binding sites? BSA 3% 30 minutes Instead, use 2% normal rabbit serum at room temperature to block the non-specific binding sites. 7. Secondary antibody ? What secondary antibody are you using? Specification (in which species was it raised against)? At what dilution(s) have you tested this antibody? Incubation, wash steps? Do you know whether the problems you are experiencing come from the secondary? What detection method are you using? Antigoat secondary antibody 1:400. Try to apply an ABC signal enhancing step. 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.

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1-10 of 15 Abreviews or Q&A

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