Anti-LAMP1 antibody (ab24170)

Thanks for the discount code. Will this be valid if I use a new lab discount on the ab24170 order?

ANSWER:

Yes, you can use this discount code to receive a free antibody if the initial vial of ab24170 is ordered using your new lab discount.

Please note however that only 1 discount code can be used per order. So when you place the order for the free antibody using the 100% discount promotional code, please make sure that the free antibody is theonly item on that order.

I hope that makes sense. Please let me know if you have any further questions.

Yes, thanks, it's very helpful to know why my "positive control" didn't work!
Thanks for sharing the location of the peptide. I would like to continue testing this
antibody to determine whether it is cross-reactive against feline LAMP1, so I will need to
order the antibody. I have a feline recombinant LAMP1 as well as feline cells to use in IP/WB,
and I can use 293T as the positive control. Could I get a promotion code for the 100% testing discount?

ANSWER:

I am very pleased to hear you would like to accept our offer and test ab24170 in feline. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for feline and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Inquiry: Is this Ab likely to recognize the recombinant human LAMP1 from R&D Systems (4800-LM, aa29-382)? I have two 7µl aliquots of this Ab (stored at -80and used one in WB, and did not see the expected bands. Since I have just one aliquot left, I'd like to know what conditions are recommended for this antibody. I loaded 62.5, 125, and 200ng/well (Bio-Rad Mini-Protean TGX 7.5%) and transferred to PVDF using the Trans-Blot Turbo 7min protocol. I also loaded a few wells with 293T lysates (5 or 15µl, 1% Triton X-100 lysis buffer from Miltenyi IP kit 130-091-124, 10^6 cells/ml) however, I had to disassemble the gel apparatus to remove the tape from the bottom of the wells and it looks like some of the MWM washed over into the lysate wells, so I can't tell whether any of the faint bands are LAMP1. I used Pierce Fast Western kit (35050) and followed the manufacturer's protocol, with 7µg of this primary Ab in 10ml diluent, for 30min at room temp, and HRP detection reagent for 10min. Detection was with Luminata substrate and ChemiDoc, 60sec exposure - no additional bands appeared with a longer exposure. I saw no LAMP1 bands when using this antibody. On a separate blot (same conditions except the gel was AnyKd instead of 7.5%), I used Millipore anti-human LAMP1 AB2971, also a rabbit polyclonal, and saw dark LAMP1 bands at the expected size (just over 100kDa), so I think my recombinant protein is OK.

ANSWER:

Thank you for your enquiry.

The immunogen peptide used to produce ab24170 corresponds to amino acids408-417 of human sequence. Thisregion would not be present in the recombinantLAMP1 from R and D systems (4800-LM, aa29-382).

I hope this is helpful. Please contact me again if you have any further questions.

Hi xxxx,

I have repeated the Lamp1 Wb with your recommended changes and have attached the data I obtained when analysing CLF enriched from HepG2 cells.

The western worked perfectly with clear bands running at ˜110 kDa, as well as bands at 45 kDa (non-glycosylated form) and ˜35 and 23 kDA (which other groups have also detected), at all 3 protein concentrations (10, 30 and 60 µg/well) and in an undiluted (Und) sample also.

Thank you again for all your help and for kindly sending more Lamp1 antibody, it was greatly appreciated.

Warm regards

ANSWER:

That does indeed look much better. Well done!

And thank you for letting me know.

If there is anything further I can help you with please do let me know, otherwise I wish you all the best with your research.

Please suggest fluoresent conjugated secondaries for:
ab48394
ab14705
ab24170
ab11420
ab2861
Using in IHC-Fr on confocal microscopy, mouse samples.
Suggested positive controls

ANSWER:

Thank you for your telephone call yesterday afternoon.

I am pleased to provide some information regarding suitable secondaries for the primary antibodies you have selected:

ab48394 LC3B antibody, ab24170 LAMP1antibody andab11420 ERp29 antibody are all rabbit polyclonal. Polyclonal antibodies contain mainly IgG isotypes, soyou need toselect an anti Rabbit IgG tested in IHC-Fr. I have selected some FITC conjugated anti rabbit IgG antibodies that are tested in IHC-Fr. I can suggest to review the online datasheets for further information. Please check that your confocal microscope will detect FITC. Also, if you require any different conjugates, please let me know and I can provide a further list for you as we have many available.

ab6717 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC)
Tested in: Flow Cyt, IHC-P, ELISA, IM, ICC/IF, IHC-Fr
http://www.abcam.com/index.html?datasheet=6717 (or use the following: http://www.abcam.com/index.html?datasheet=6717).

ab7086 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC), pre-adsorbed
Tested in: Flow Cyt, IHC-P, ELISA, IM, ICC/IF, IHC-Fr
http://www.abcam.com/index.html?datasheet=7086 (or use the following: http://www.abcam.com/index.html?datasheet=7086).


The ab14705 MTCO1 antibody [1D6E1A8] and ab2861 SERCA2 ATPase antibody [2A7-A1] are both mouse monoclonal IgG2a.

We have no anti mouse IgG2a secondaries tested in IHC-Fr, but in this case you canuse a general anti-mouse IgG.I have selected an FITC conjugated secondary, but again please let me know if you would prefer other conjugates as we have many available.

ab6785 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC)
Tested in: IHC (PFA fixed), Flow Cyt, IHC-P, WB, ELISA, IM, ICC/IF, IHC-Fr
http://www.abcam.com/index.html?datasheet=6785 (or use the following: http://www.abcam.com/index.html?datasheet=6785).

Both ab2861 and ab14705 are mouse antibodies. These are tested successfully in mouse, and so I would like to reassure you we can provide our guarantee for the results. However, because you are staining mouse tissue, I can suggest it may be better to select primary antibodies that are not raised in mouse. When you add the anti-mouse secondary during the staining, it is possible this can bind to mouse antibodies within the tissue and give some background staining.

You may like to consider the following instead:
ab3625 SERCA2 ATPase antibody
Rabbit polyclonal
Tested in: IHC-P, IP, WB, ICC/IF, IHC-Fr
Reacts with: Hu, Ms, Rat, Pig, Rb

I am sorry there are no alternatives for the MTCO1.

If you require some further information regarding antibodies and IHC, as discussed on the telephone I am including some links from our website. These can be found from the scientific support tab on the homepage:

Antibody guide:
http://www.abcam.com/index.html?pageconfig=resource&rid=11287

Protocols (Select Immunostaining from the list on the following protocolspage):
http://www.abcam.com/index.html?pageconfig=popular_protocols

Fluorochrome guides:
http://docs.abcam.com/pdf/immunology/fluorochrome_guide.pdf
http://www.abcam.com/index.html?pageconfig=resource&rid=11755

How to choose the right secondary:
http://www.abcam.com/index.html?pageconfig=resource&rid=12825

How to choose a conjugate:
http://www.abcam.com/index.html?pageconfig=resource&rid=12922

And the following external website may also be helpful:
http://www.ihcworld.com/

I hope this will be helpful. If you require any further information or assistance, please do not hesitate to contact us (mailto:technical@abcam.com).