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  1. Link

    m6pr-cation-independent-antibody-ab32815.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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Anti-M6PR (cation independent) antibody (ab32815)

  • Datasheet
Reviews (6)Q&A (3)References (25)

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Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody (ab32815)
  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)

Key features and details

  • Rabbit polyclonal to M6PR (cation independent)
  • Suitable for: ICC/IF, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-M6PR (cation independent) antibody
    See all M6PR (cation independent) primary antibodies
  • Description

    Rabbit polyclonal to M6PR (cation independent)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Full length native protein (purified) corresponding to Cow M6PR (cation independent). Full length native protein purified from adult bovine liver tissue.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: Whole serum
  • Concentration information loading...
  • Purity

    Whole antiserum
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Endosome
    • Signal Transduction
    • Protein Trafficking
    • Golgi Proteins
    • Neuroscience
    • Processes

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32815 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
ICC/IF (2)
1/50 - 1/500.
IHC-P
1/1000.
Notes
ICC/IF
1/50 - 1/500.
IHC-P
1/1000.

Target

  • Function

    Transport of phosphorylated lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. Lysosomal enzymes bearing phosphomannosyl residues bind specifically to mannose-6-phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex. This receptor also binds IGF2. Acts as a positive regulator of T-cell coactivation, by binding DPP4.
  • Sequence similarities

    Belongs to the MRL1/IGF2R family.
    Contains 1 fibronectin type-II domain.
  • Domain

    Contains 15 repeating units of approximately 147 AA harboring four disulfide bonds each. The most highly conserved region within the repeat consists of a stretch of 13 AA that contains cysteines at both ends.
  • Cellular localization

    Lysosome membrane. Colocalized with DPP4 in internalized cytoplasmic vesicles adjacent to the cell surface.
  • Target information above from: UniProt accession P11717 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3482 Human
    • Entrez Gene: 16004 Mouse
    • Omim: 147280 Human
    • SwissProt: P11717 Human
    • SwissProt: Q07113 Mouse
    • Unigene: 487062 Human
    • Unigene: 673278 Human
    • Unigene: 26553 Mouse
    • Unigene: 486511 Mouse
    see all
  • Alternative names

    • 300 kDa mannose 6 phosphate receptor antibody
    • 300 kDa mannose 6-phosphate receptor antibody
    • Cation independent mannose 6 phosphate receptor antibody
    • Cation-independent mannose-6-phosphate receptor antibody
    • CD222 antibody
    • CD222 antigen antibody
    • CI Man 6 P receptor antibody
    • CI Man-6-P receptor antibody
    • CI MPR antibody
    • CI-M6PR antibody
    • CI-MPR antibody
    • CIMPR antibody
    • IGF 2 receptor antibody
    • IGF 2R antibody
    • IGF II receptor antibody
    • IGF-II receptor antibody
    • IGF2 receptor antibody
    • Igf2r antibody
    • Insulin like growth factor 2 receptor antibody
    • Insulin like growth factor II receptor antibody
    • Insulin-like growth factor 2 receptor antibody
    • Insulin-like growth factor II receptor antibody
    • M6P R antibody
    • M6P/IGF2 receptor antibody
    • M6P/IGF2R antibody
    • M6PR antibody
    • mannose 6 phosphate receptor antibody
    • mannose 6 phosphate receptor, cation independent antibody
    • MPR 300 antibody
    • MPR300 antibody
    • MPRI antibody
    • MPRI_HUMAN antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
    Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)

    Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent) (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
    Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)

    Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent)(green) showing staining in the cytoplasm and nucleus of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
    Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)

    Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent)  (green) showing staining in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody (ab32815)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody (ab32815)
    Ab32815 staining human normal left ventricle of heart. Staining is localized to lysosome and lysosomal membrane.
    Left panel: with primary antibody duluted at 1:1000. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplifi
  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
    Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody (ab32815)
    ICC/IF image of ab32815 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32815, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (25)

Publishing research using ab32815? Please let us know so that we can cite the reference in this datasheet.

ab32815 has been referenced in 25 publications.

  • Romano R  et al. Alteration of the late endocytic pathway in Charcot-Marie-Tooth type 2B disease. Cell Mol Life Sci 78:351-372 (2021). PubMed: 32280996
  • Haldar S  et al. Precise Triggering and Chemical Control of Single-Virus Fusion within Endosomes. J Virol 95:N/A (2020). PubMed: 33115879
  • Margiotta A  et al. Invariant chain regulates endosomal fusion and maturation through an interaction with the SNARE Vti1b. J Cell Sci 133:N/A (2020). PubMed: 32907852
  • Finetti F  et al. The intraflagellar transport protein IFT20 controls lysosome biogenesis by regulating the post-Golgi transport of acid hydrolases. Cell Death Differ 27:310-328 (2020). PubMed: 31142807
  • Min SH  et al. PIKfyve Deficiency in Myeloid Cells Impairs Lysosomal Homeostasis in Macrophages and Promotes Systemic Inflammation in Mice. Mol Cell Biol 39:N/A (2019). PubMed: 31427458
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Western blot abreview for Anti-M6PR (cation independent) antibody - Lysosome Membrane Marker

Average
Abreviews
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (293T)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted May 08 2019

Western blot abreview for Anti-M6PR (cation independent) antibody

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Sheep Tissue lysate - whole (Heart)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
75 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More

Abcam user community

Verified customer

Submitted Aug 09 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-M6PR (cation independent) antibody - Lysosome Membrane Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (primary alveolar macrophage)
Permeabilization
Yes - TX-100
Specification
primary alveolar macrophage
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Feb 05 2009

Western blot abreview for Anti-M6PR (cation independent) antibody - Lysosome Membrane Marker

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - other (Membrane fraction of Hela Cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
100000 cells
Specification
Membrane fraction of Hela Cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Sep 12 2008

Immunocytochemistry/ Immunofluorescence abreview for Anti-M6PR (cation independent) antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela Cells)
Permeabilization
No
Specification
Hela Cells
Fixative
Methanol
Read More

Abcam user community

Verified customer

Submitted Aug 11 2008

Western blot abreview for Anti-M6PR (cation independent) antibody - Lysosome Membrane Marker

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (lacrimal gland)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
lacrimal gland
Blocking step
Rockland Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Read More

Abcam user community

Verified customer

Submitted Jul 10 2008

Question

I am trying to work the problem out in a different way. To that end, pl. don’t send me a replacement vial but pl. issue a credit memo for the same. I will place an order for the ab32815 and use it with a secondary conjugated to a flurochrome. 32815 has worked for me, the trouble is I have not be able to find a replacement for it that has a flurochrome attached and works as efficiently as 32815.

Read More

Abcam community

Verified customer

Asked on Nov 28 2012

Answer

Thank you for your reply.

If you would prefer, I would be happy to issue ab32815 as a free of charge replacement for ab129923.

I look forward to your reply. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Abcam Scientific Support

Answered on Nov 28 2012

Question

Staining of cells is diffuse in the cytoplasm and there is heavy background outside the cells.

Read More

Abcam community

Verified customer

Asked on Aug 13 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. The one protocol modification that might have an effect on the background stain would be to substitute either a different serum or BSA (1%) for the blocking donkey serum, but if you have not seen this issue with other antibodies in the same protocol with the same reagents, I am doubtful the susbstitution will help.

I have issued a free of charge replacement with the order number 1144831.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

Read More

Abcam Scientific Support

Answered on Aug 13 2012

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific band, and high background SAMPLE Cell extract PRIMARY ANTIBODY Abacm ab32815, tried 1:1000, 1:500 in TBST with 1%milk cold room, overnight DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED we have the cells tested by RT-PCR for Igf2r expression, also have knock-out cells as negative which were confirmed by RT-PCR ANTIBODY STORAGE CONDITIONS -20 C after receive it SAMPLE PREPARATION RIPA buffer lysis, plus protease inhibitors (Pierce Cat 78410) AMOUNT OF PROTEIN LOADED 25 ug per line ELECTROPHORESIS/GEL CONDITIONS Pierce Precise protein gel (4-20%) TRANSFER AND BLOCKING CONDITIONS Tris-Glycine transfer buffer, Blocking in TBST with 5% milk, room tempeture for 1.5hr SECONDARY ANTIBODY Jackson ImmunoResearch 711-035-152 1:10,000 dilution work well in the lab HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? last step after got the results ADDITIONAL NOTES many unspecific bands on the film, no difference between positive and negative samples PO number for this: RSTFD0000209430

Read More

Abcam community

Verified customer

Asked on Sep 04 2007

Answer

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab32815 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. What species of cell extract are you using? In most cases, the cause of multiple bands is because the primary and/or secondary antibody concentration is too high and causes non-specific binding. The dilution that we have on the datasheet is a recommended starting dilution and customers are encouraged to determine the optimal concentration/dilution. You can try decreasing the primary (1:2000 ~ 1:5000). Also, running a no-primary control experiment will be able to eliminate the possibility that your secondary antibody is binding non-specifically. What are the band sizes you obtained? The bands located above the expected molecular weight band could be multimers. Please try boiling your protein in SDS-PAGE for 10 minutes to ensure proper disruption of multimers. This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody. Meanwhile, bands located below the expected molecular weight may indicate that your target protein has been digested. Please make sure that you incorporated sufficient protease inhibitors and have kept your samples on ice the whole time. If you can provide an image, that would certainly help me understand the problem better. Since you mentioned that you seeing high background, I would assume that it is the milk that is causing the problem. In general, there are two blocking solutions that are widely used, non-fat milk and BSA. We would recommend using 5% BSA as almost all of our products are tested with it and have proven to be effective. Most customers who use milk usually have high background, due to the secondary antibody binding to the milk. You can check if this dark signal you are detecting is from the secondary or not by incubating your blot in secondary only. Increasing the washing frequency and duration would also help eliminate high background. Can you confirm that you have used Tween in your washing solution and have washed at least 15min x 3 times? Please try this if you have not already done so. I am sorry for the string of questions but I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me with the answers to the above and your shipping address. Also, please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you.

Read More

Abcam Scientific Support

Answered on Sep 04 2007

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