Overview

  • Product name
  • Description
    Rabbit polyclonal to MCP1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant full length protein (Rat).

  • Positive control
    • RAW 264.7 cells (mouse macrophage/monocyte adherent cell line) stimulated with LPS for 15 hours

Properties

Applications

Our Abpromise guarantee covers the use of ab7202 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-P 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration. PubMed: 21088249
ICC/IF Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 18 kDa.

Target

  • Function
    Chemotactic factor that attracts monocytes and basophils but not neutrophils or eosinophils. Augments monocyte anti-tumor activity. Has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis or atherosclerosis. May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.
  • Sequence similarities
    Belongs to the intercrine beta (chemokine CC) family.
  • Post-translational
    modifications
    Processing at the N-terminus can regulate receptor and target cell selectivity. Deletion of the N-terminal residue converts it from an activator of basophil to an eosinophil chemoattractant.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • C-C motif chemokine 2 antibody
    • CCL2 antibody
    • CCL2_HUMAN antibody
    • Chemokine (C C motif) ligand 2 antibody
    • GDCF 2 antibody
    • GDCF-2 antibody
    • GDCF2 antibody
    • HC11 antibody
    • HSMCR30 antibody
    • JE antibody
    • MCAF antibody
    • MCP 1 antibody
    • MCP-1 antibody
    • MCP1 antibody
    • MGC9434 antibody
    • Monocyte chemoattractant protein 1 antibody
    • Monocyte chemotactic and activating factor antibody
    • Monocyte chemotactic protein 1 antibody
    • Monocyte secretory protein JE antibody
    • SCYA2 antibody
    • Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig je) antibody
    • Small inducible cytokine A2 antibody
    • Small inducible cytokine subfamily A (Cys Cys), member 2 antibody
    • Small-inducible cytokine A2 antibody
    • SMC CF antibody
    • SMC-CF antibody
    • SMCCF antibody
    see all

Images

References

This product has been referenced in:
  • Wang F  et al. Myeloid ß-Catenin Deficiency Exacerbates Atherosclerosis in Low-Density Lipoprotein Receptor-Deficient Mice. Arterioscler Thromb Vasc Biol 38:1468-1478 (2018). Read more (PubMed: 29724817) »
  • Xue XT  et al. Sexual dimorphism of estrogen-sensitized synoviocytes contributes to gender difference in temporomandibular joint osteoarthritis. Oral Dis N/A:N/A (2018). Read more (PubMed: 29806726) »
See all 54 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (liver)
Loading amount
30 µg
Specification
liver
Treatment
2mg/kg LPS 1h
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 16 2012

Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Answer

Thank you for your note.



This is to let you know that I have just contacted and asked our Account Department to raise a credit note for you - for the cost of one vial of ab7202. For your information, the internal reference note for this credit is CN20561. You can use this credit in the near future for any of the products which are in the catalogue.



I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question

Dear abcam technical support team:

This customer has purchase ab7202 (Anti-MCP1 antibody) and has conducted the WB several times with mouse sample.

The results show no band; moreover, this customer also has used RAW 264.7 cells and stimulated with LPS as a positive control, but the results also shows no band,

she thinks our antibody was lost its function already. Therefore, she would like to ask for your help to solve her problem,

I also attached her data and experiment step with different antibody in this letter, please check this and could you please offer any help to this customer,

thanks for your assistance. Best regards



1. Order details:


Batch number: Lot: GR47175-3

Abcam product code: ab7202

Antibody storage conditions (temperature/reconstitution etc) : -20 ℃ freezer




2. Please describe the problem (high background, wrong band size, more bands, no band etc).

NO band

3. On what material are you testing the antibody in WB?


Species: mouse

What’s cell line or tissue: renal tissue

Cell extract or Nuclear extract: cell extract

Purified protein or Recombinant protein: purified protein




3. The lysate


How much protein was loaded: 150 μg

What lysis buffer was used: RIPA



What protease inhibitors were used: protease inhibitor cocktail, PMSF, NaF NaVO3

What loading buffer was used: SDS sample buffer with protein dye

Phosphatase inhibitors: none

Did you heat the samples: temperature and time: 100℃, 5 min




4. Electrophoresis/Gel conditions/ Transfer conditions


Reducing or non reducing gel: reducing gel

Reducing agent: 2-mercaptoethanol

Gel percentage : 12%

Transfer conditions: (Type of membrane, Protein transfer verified): PVDF membrane, protein transfer apparatus from ATTO corporation (Model: AE6675)




5. Blocking conditions


Buffer: PBS with 0.1 % Tween 20

Blocking agent: milk, BSA, serum, what percentage: 3% BSA

Incubation time: 60 min

Incubation temperature: 25 ℃




6. Primary Antibody


Species: rabbit

Reacts against: MCP-1



At what dilution(s) have you tested this antibody: 1:1000

What dilution buffer was used: 3% BSA in PBS with 0.1% of Tween 20

Incubation time: 18 hours

Incubation temperature: 4 ℃

What washing steps were done: washing membrane with PBS with 0.1% of Tween 20 for 6 times, 5 minutes per each time




7. Secondary Antibody


Species: goat

Reacts against: rabbit antibody

At what dilution(s) have you tested this antibody: 1:3000

Incubation time: 1 hour

Wash steps: washing membrane with PBS with 0.1% of Tween 20 for 6 times, 5 minutes per each time

Fluorochrome or enzyme conjugate: HRP conjugate

Do you know whether the problems you are experiencing come from the secondary? NO




8. Detection method
ECl, ECl+, other detection method: ECL



9. Did you apply positive and negative controls along with the samples? Please specify. Yes, the cell lysate of RAW264.7

10. Optimization attempts


How many times have you tried the Western? 2 times

Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): NO

Do you obtain the same results every time e.g. are background bands always in the same place? NO

What steps have you altered? NO

Read More
Answer

Thank you for your enquiry regarding ab7202 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this antibody.

The protocol looks fine to me. It may well be the antibody has been accidentally damaged.

I can offer a new vial free of charge for your troubled customer but all our vials are from the same belled and purification. Could you please confirm the Abcam Order and the date of purchase and let me know how your customer would like to proceed with this enquiry.

I look forward to hearing from you soon.

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Question
Answer

Thank you for your phone call today, and I apologize for any miscommunication with my colleague yesterday. Your credit note ID for ab20641 is: ***. Your credit note ID for ab7202 is: ***. I am sorry that these antibodies did not perform as stated on the datasheet. If payment has already been made on the original orders and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.  

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Question
Answer

Thank you for your call today and for your questions about ab7202. As we discussed, we recommend using a microwave antigen retrieval method with this antibody. However, if the antibody is still staining the wrong localization, please let me know and we will be happy to send a replacement or issue a credit or refund. I look forward to hearing from you. Let me know if you have any questions.

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Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (injred spinal cord tissue)
Specification
injred spinal cord tissue
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Ms. soo ryung jeong

Verified customer

Submitted Oct 01 2009

Question

BATCH NUMBER 7052 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Western blot: no bands detectable Immunohistochemistry: no reaction SAMPLE WB: mouse aorta protein extract IHC: mouse frozen cross sections of the aorta, OCT embedded PRIMARY ANTIBODY WB: 1:2000, 1:1000, 1:500 over night, wash step 2x 5min in TBST 0.05% IH: over night 1:400, 1:200, 1:100, wash step 3x 5min in PBS SECONDARY ANTIBODY WB: Amersham donkey anti-rabbit IgG HRP, 1:3500, 60mion, wash step 1x 15min in TBST 0.05% IH: Vector/Linaris goat anti-rabbit IgG Biotin in PBS, 1:200, 45min or 60min, wash step 3x 5min PBS DETECTION METHOD WB: ECL IH: DAB-chromogen (Dako) POSITIVE AND NEGATIVE CONTROLS USED IH: positive: serial sections that showed positive staining with thsi primary asntibody, but different Lot (56165) negative: unspecific IgG SAMPLE PREPARATION WB: RIPA buffer + protease inhibitor (complete, mini EDTA-free, Roche) 10:1 loading buffer RotiLoad 1 (Roth) 1:4, 95?C, 5min TRANSFER AND BLOCKING CONDITIONS WB: transfer: transfer buffer (25mM Tris base, 0.2M glycine, 20% methanol, pH 8.5, 60min 340mA or transfer over night 90mA block: 5% milk/TBST, 1h IH: block: 6% goat serum in 4% BSA/PBS or 4% goat serum HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4both HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? WB: first antibody concentration, blocking concentration as indicated IH: first antibody concentration, incubation period, blocking agents as indicated, changed other reagents and tested them iwth another primary antibody in order to find out whether they are still working (positive results).

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Answer

I'm sorry to hear the customer is having problems with his new vial of ab7202. Thank you for taking the time to fill in the questionnaire, I can see clearly that the protocol looks very good and the antibody should perform if the customer has previously had good results. Could you please confirm the lot number or the order number for this vial as we have no record of a lot 7052 and confirm the storage conditions, my apologies for the inconvenience. Could you please also send us an image of the positive staining with lot 56165? Thank you for your patience, we look forward to hearing from you soon,

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 57806 DESCRIPTION OF THE PROBLEM low signal and high background using a biotinylated secundary antibody non-signal using a FITC-conjugated secondary antibody SAMPLE Aortic frozen sections from mice (apo E-/- and LDL-/- PRIMARY ANTIBODY Rabbit polyclonal to MCP-1 (ab7202, Abcam)incubated a)for 2-3 hours at room temperature b)overnight at 4?C Dilutions used: 1/25, 1/50, 1/100, 1/500, 1/1000 Wash step: Rinse 2x5 min in PBS SECONDARY ANTIBODY a)Biotinylated goat anti-rabbit (Vector) Incubation at room temperature for 2-3 h. Dilution used: 1/200 (in BSA 1%) b)FITC-goat anti rabbit (Alexa Fluor) Incubation at room temperature for 3 h. Dilution used: 1/300 DETECTION METHOD a)ABC complex (Vector): used in a 1/200 dilution and incubated for 45 min at room temperature.Developed with DAB chromogen at room temperature,counterstained with haematoxylin, dehydrated and mounted in DPX. b)Mounted in Mowiol. POSITIVE AND NEGATIVE CONTROLS USED Positive controls : aorta from mice (32 and 24 weeks of age) fed with a high-cholesterol diet for either 22 or 14 weeks. Negative controls: the same tissue without adding the primary antibody. ANTIBODY STORAGE CONDITIONS 4?C FIXATION OF SAMPLE Acetone for 5 min ANTIGEN RETRIEVAL None PERMEABILIZATION STEP Sodium deoxycholate for 10 min 0.5 % Triton in PBS solution for 10 min BLOCKING CONDITIONS 1%BSA in PBS for 30 min at room temperature avidin/biotin for 10 min each at room temperature. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES I would like to know whether someone has already tried this antibody in aorta from mice.

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Answer

I'm sorry to hear you are having problem with ab7102. We have not tested it in mouse aorta or have any reviews yet from customers using it in aorta. So please do let us know when your staining works and we will reward you with an Amazon gift voucher (a review can be submitted via our online datasheet). I would like to suggest the following modifications to your protocol: -wash between all steps minimum 4 times 5min in PBS -fix for 10min in methanol or acetone pre-cooled in the freezer -block endogenous peroxidases with hydrogen peroxide (0.3% v/v in PBST) -block the slides with normal goat serum 10% v/v 1hr -use PBST (0.1-0.5%v/v triton) to dilute the serum, the primary and secondary antibody. -Incubate the biotin antibody for 2 hours at 1:400 in PBST, no BSA. -the dilution of the detection kit from Vector can be altered to decrease the background, make sure you prepare the ABC in PBS and leave to rest for 30min before applying onto slides. Try 1:500 for 1hr. It is best to leave it longer but more dilute. -When washing the slides after the DAB reaction, use Tris buffer 0.5M rather than PBS. This will decrease background as well. If you want to use the FITC-conjugated antibody dilute it in PBST (pH 7 or 8). For background problems please always check the secondary antibodies are not responsible. To do so, do the same protocol simply omitting the primary antibody. Please let me know if this helps and do not hesitate to contact us for further advice,

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Question
Answer

Thank you for your phone call. There are no special conditions that need to be followed to use ab7202 in IHC-frozen sections and so I suggest fixing your sections with acetone as per our general IHC protocol for frozen sections (the protocol can be found in the protocols tab of the online datasheet).

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1-10 of 16 Abreviews or Q&A

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