Overview

  • Product name

    Mouse SDF1 ELISA Kit
    See all SDF1 alpha kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 3 pg/ml
  • Range

    9.38 pg/ml - 600 pg/ml
  • Recovery

    99 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 99.34 89% - 107%
    Serum 99 91% - 107%
    Plasma 100.36 90% - 107%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Mouse SDF1 ELISA (Enzyme-Linked Immunosorbent Assay) kit (ab100741) is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse SDF1 in serum, plasma and cell culture supernatants. (Mouse SDF1 concentration is pretty low in normal serum/plasma, it may not be detected in this assay).


    This assay employs an antibody specific for mouse SDF1 coated on a 96-well plate. Standards and samples are pipetted into the wells and SDF1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse SDF1antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of SDF1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Mouse SDF1 (lyophilized) 2 vials
    Recombinant Mouse SDF1 Standard (lyophilized) 2 vials
    SDF1 Microplate (12 strips x 8 wells) 1 unit
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Relevance

    SDF1 alpha and 1 beta are small cytokines that belong to the intercrine family, members of which activate leukocytes and are often induced by proinflammatory stimuli such as lipopolysaccharide, TNF or IL1. SDF1 alpha has been shown to be chemotactic for lymphocytes and is recently reported to be a ligand for CXCR4 (LESTR/fusin), a co-receptor for HIV1 entry into T cells. SDF-1 binding to CXCR4 inhibits HIV1 entry.
  • Alternative names

    • C-X-C motif chemokine 12
    • Chemokine (C X C motif) ligand 12
    • CXCL12
    • hIRH
    • Intercrine reduced in hepatomas
    • IRH
    • PBSF
    • Pre B cell growth stimulating factor
    • SCYB12
    • SDF 1
    • SDF 1 beta
    • SDF 1b
    • SDF1
    • SDF1 alpha (3-67)
    • SDF1 beta (3-72)
    • SDF1A
    • SDF1B
    • Stromal cell derived factor 1
    • TLSF a
    • TLSF b
    • TLSFa
    • TLSFb
    • TPAR1
    see all

Applications

Our Abpromise guarantee covers the use of ab100741 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SDF1 measured in culture supernatants from control or activated cells (P/L), stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297) with the addition of 1 ug x mL-1 of LPS (Sigma) for the last 6 hours, and in biological fluids (duplicates +/- SD). All samples tested undiluted.

  • Standard curve with background signal subtracted (duplicates; +/- SD).

  • Representative Standard Curve using ab100741

  • Representative Standard Curve using ab100741

Protocols

References

This product has been referenced in:

  • Xu S  et al. CXCR7 promotes melanoma tumorigenesis via Src kinase signaling. Cell Death Dis 10:191 (2019). Read more (PubMed: 30804329) »
  • Li X  et al. Transplantation of skin mesenchymal stem cells attenuated AngII-induced hypertension and vascular injury. Biochem Biophys Res Commun 497:1068-1075 (2018). Read more (PubMed: 29481801) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Mouse SDF-1 ELISA

Good Excellent 5/5 (Ease of Use)
Abreviews
I used the Mouse SDF-1 ELISA kit to check the levels in mouse retinal tissues and it worked.

Dr. Finny Monickaraj

Verified customer

Submitted Aug 19 2015

Answer

Both antibodies used in this kit recognize a region within Lys22­ - Lys89 (Accession # Q6ICW0). Thus, although not previously tested, we do not expect an additional sequenced fused to the C-terminus to interfere with detection.

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Answer

Thank you for your reply.

You may want to try the new lots and take the precautions listed in the previous email. The lab does not think that anything is wrong with the standards as all lots are quality control tested. Just take the necessary steps and as long as you get the standard curve, you should still be able to use it with your samples since most of the samples will probably fall within the range you're detecting.

Plus, why is the blank so high? You're seeing a reading similar to 37.5 pg/ml. Do you have any reasoning for that?

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your inquiry and your patience while I looked into this further.

The lab has QC'ed these standards for the kit and the certificate of analysis for a recent batch is attached.

The150 rpm is a little fast but it is not likely that your rocking/shaking speed is causing the weak standard signals. The purpose of rocking is simply to ensure that the sample and reagents completely and evenly cover the coated antibody wells. As long as the solutions in the wells are not spilling over to other wells andyou are rocking gently, ˜0.5-1 cycles/sec, that is fine. We use the roto-shake genie from Scientific Industries if that helps. Please note that all incubations should be performed under gentle rocking.

Below are some general tips for ensuring a strong signal strength. Keep in mind that it is perfectly fine if your resulting standard signals are a little higher or lower than this. In fact, if you graph your standards under "New std x2," you get an r^2 value of 0.977, even including the blank which is good. Youcould consider doing the optional overnight sample/standard incubation (if not doing so already) to help boost overall signal strength though.

Please note the following precautions to ensure a stronger OD response from your standard:

1. When preparing your standards, it is very critical to briefly spin down the vial first. The powder may drop off from the cap when opening it if you do not spin down. Be sure to dissolve the powder thoroughly when reconstituting. After adding Assay Diluent to the vial, we recommend inverting the tube a few times, then flick the tube a few times, and then spin it down; repeat this procedure 3-4 times. This is a technique we find very effective for thoroughly mixing the standard without too much mechanical force.

2. Do not vortex the standard during reconstitution, as this will destabilize the protein.

3. Once your standard has been reconstituted, it should be used right away or else frozen for later use.

4. Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature.

5. Be sure to discard the working standard dilutions after use – they do not store well.

Lastly, there is an extra vial of standard (Item C, 2 vials total) to minimize storage issues. Using a fresh, unreconstituted Item C vial is always prefereable to a used/stored reconstituted vial. Ifyou must re-use the reconstituted standard (the 9 ng/ml solution) it should be stored at -80ºC for only 1-2 weeks maximum. The diluted standards should not be stored.

I hope this information helps. If you have any other questions, please call 888-772-2226.

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Answer

Thank you for your reply.

Just to clarify, the diluted/working standards (600, 200, 66.7, 22.2, 7.4, 2.5, 0.82 pg/ml) should not be stored. The reconstituted standard (50 ng/ml) can be stored at -80ºC for 1-2 weeks at most. Remember, an extra vial of standard (Item C) is included in each kit (2 vials total) to minimize storage issues.

The example above is for theIL6 mouse ELISAkit but the same storage guidelines apply to the SDF-1a kit.
I hope this information helps. Please contact us with any other questions.

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Answer

Thank you very much for your email.

I am sorry to hear about the customer who has decided to pull out from getting this product. We have helped your customer as far as we can by providing them credit note which you have used for order 112737. I have checked that spinchem.cz is not an official distributor of Abcam so the Abpromiose guarantee will not apply. This is because we cannot control the product shipping conditions when our products are resold by unofficial distributor; these products are very temperature sensitive so we can not quality check how the unofficial distributor send these products to their customers. We have official distributor in Czeck republic, please tell your customer to buy the product from them.

I am sorry we will be unable to recall the product this because Abpromise guarantee does not applies spinchem.cz.

I hope this information will nevertheless be helpful. Should you require more information please do not hesitate to contact me.

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Answer

Thank you for your inquiry.

No, the protocol is right. Steps 4 and 5 are not the same as steps 6 and 7. Steps 4 and 5 involve the biotinylated antibody and steps 6 and 7 involved the streptavidin-HRP solution.

Yes, step 6 in "Assay Method"is the streptavidin-HRP from "Preparation of Reagents" step 7.

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your inquiry.

We know of the IL6 ELISA kit being tested in liver tissue lysate, but unfortunately we do not have any info on brain lysate.

For the SDF1 alpha ELISA kit, we haven't tested tissues with this kit but it would most likely work.If you decide to test this kit with mouse brain tissue lysates samples, we recommend diluting the samples at least 5-fold with Assay Diluent B to minimize any effects of the detergents in the lysis buffer. The samples may need to be diluted further but this would need to be determined empirically.

If you would like to test ab100741 in tissues, we do offer a testing discount program. Essentially you would purchase the kit up front at cost, test it in tissues, and submit an Abreview letting us know whether or not it works. You would then receive a discount code worth the purchase price (currently USD$470.00) off your next Abcam order within 4 months.

If you decide to test in tissues, here are some protocol recommendations.For the original lysate, in general you would want to aim for at least 1 mg/ml protein, preferably more, to achieve 50-500 ug/ml after dilution. Again though, this range should just be used as a starting point and the optimal concentration would need to be determined empirically.
In brief, a lysis buffer must meet the following specifications:
A) has relatively low salt content (700 mM or less)
B) does not contain sodium azide
C) does not contain >0.1% SDS
D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)
This would include any buffers used for immunoprecipitations, including RIPA buffer.
After homogenization, spin down the lysates to remove cell/tissue debris (5 min @ 10000 x g or 10 min @ 5000 x g) and save the supernatant. Lysates should be frozen as soon as possible, and stored at -20°C (or -80°C, if possible). Spin them again before incubating with the antibody array. Determine the protein concentration of your lysates (for example the bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay. Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 uL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target for total protein concentration of the homogenate should be at least 1000 ug/mL, but 2000 ug/mL would be better.
I hope this information helps. Please contact us with any other questions.

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Answer

After reviewing the standard OD data, it doesn’t look like there is any significant detection or linear pattern. Did you run any samples as this would help determine if the issue is just with the standard protein or a common detection component? Strictly based of the standard data available here, it looks like the highest OD measured (0.096) was from the 0 pg/ml which is surprising as this should be just straight diluent and therefore give back the lowest absorbance reading. However, it is entirely possible that the standard ODs values, including the 0 standard, are not real but just back noise. If you have any microplate strip left, I would advise you to try and repeat the experiment keeping the following tips in mind, if you run samples as well and those do not have any detectable signals this would point to a common detection component being the issue. Detection Antibody – once reconstituted, this must be used within a couple of days. We provide 2 tubes, enough for half a plate each. Be sure to centrifuge before reconstitution. HRP-Streptavidin – comes as a liquid concentrate. Before withdrawing any liquid from the tube, be sure to centrifuge the vial and pipet up and down to mix thoroughly, as precipitation may form in storage. Once diluted, the HRP-strep must be used up that day. Do not store. Also do not dilute with Assay Diluent A as it contains sodium azide which inhibits HRP activity. TMB Solution - This is stable for 6 months at 4 degrees C, but it is light-sensitive. Pre-coated ELISA plate - once the microplate wells have been opened, they must be stored at 2-8oC and used within one month. Return unused wells to the pouch containing desiccant pack, and reseal along the entire edge. The entire kit should be stored at 4 degrees C (for up to 6 months from date of shipment) or -20 degrees C for longer storage. Avoid multiple freeze-thaw cycles. If after running the samples there were detectable sample signals, then this indicates the issue is most likely with the standard protein only and you should keep the following tips in mind for ensuring proper standard signal strength. When preparing your standards, it is very critical to briefly spin down the vial first. The powder may drop off from the cap when opening it if you do not spin down. Be sure to dissolve the powder thoroughly when reconstituting. After adding Assay Diluent to the vial, we recommend inverting the tube a few times, then flick the tube a few times, and then spin it down; repeat this procedure 3-4 times. This is a technique we find very effective for thoroughly mixing the standard without too much mechanical force. Do not vortex the standard during reconstitution, as this will destabilize the protein. Once your standard has been reconstituted, it should be used right away or else frozen for later use. Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature. Be sure to discard the working standard dilutions after use – they do not store well. And keep in mind that that we provide 2 vials of lyophilized standard in the kit, to minimize storage issues. The unopened vials will be stable at 4 degrees for several months. I hope you find this information useful. Please let me know if you have any questions or comments.

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Answer

Thank you for contacting us. I am happy to offer you some tips to help you improve better results from this kit. I would also appreciate if you could send me the raw OD data, so we can try to find out why the standards provided such low values. In the meantime, I would suggest following these tips, to obtain a stronger OD response from the standards: -When preparing your standards, it is very critical to spin down the vial first. The powder may drop off from the cap when opening it if you do not spin down. Be sure to dissolve the powder thoroughly when reconstituting. -After adding Assay Diluent to the vial, we recommend inverting the tube a few times, then flick the tube a few times, and then spin it down; repeat this procedure 3-4 times.  This is a technique we find very effective for thoroughly mixing the standard without too much mechanical force. -Do not vortex the standard during reconstitution, as this will destabilize the protein. Once your standard has been reconstituted, it should be used right away or else frozen for later use. -Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature.  -Be sure to discard the working standard dilutions after use – they will not store well.    And keep in mind that that we provide 2 vials of lyophilized standard in the kit, to minimize storage issues I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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