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Recombinant Anti-M6PR (cation independent) antibody [EPR6599] KO Tested (ab124767)

Validated using a knockout cell line

Recombinant

RabMAb

Recombinant Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

Western blot - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Western blot - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Western blot - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Western blot - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • OI-RD Scanning - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

  • Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6599] to M6PR (cation independent)
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405

Alexa Fluor® 555

Alexa Fluor® 594

Alexa Fluor® 647

APC

Carrier Free

PE

<!–

Overview

  • Product name

    Anti-M6PR (cation independent) antibody [EPR6599]

    See all M6PR (cation independent) primary antibodies

  • Description

    Rabbit monoclonal [EPR6599] to M6PR (cation independent)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF

    more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human M6PR (cation independent) aa 2450 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link:

    P11717

    (Peptide available as

    ab169957

    )

  • Xem thêm: Quy tắc thêm s, es và cách phát âm chuẩn đuôi s,es trong tiếng anh

    Positive control

    • Jurkat, 293T, C6, PC-12, NIH/3T3 and Caco-2 cell lysates; Human papillary carcinoma, thyroid and tonsil tissue; Mouse heart, kidney, colon and spleen tissue; Rat colon tissue . ICC/IF: HAP1 WT and HAP1-IGF2R knockout cells
  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • – High batch-to-batch consistency and reproducibility
    • – Improved sensitivity and specificity
    • – Long-term security of supply
    • – Animal-free production

    For more information

    see here

    .

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to

    RabMAb® patents

    .

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 3.90 x 10 -12 M

    Learn more about KD

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading…
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6599
  • Isotype

    IgG
  • Research areas

    • Immunology

    • Innate Immunity

    • Macrophage / Inflamm.

    • Tags & Cell Markers

    • Subcellular Markers

    • Organelles

    • Endosome

    • Signal Transduction

    • Protein Trafficking

    • Golgi Proteins

Associated products

  • Xem thêm: Một số bài toán nâng cao lớp 2 có lời giải-Baitoan.com

    Alternative Versions

    • Alexa Fluor® 488 Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab184807)

    • Alexa Fluor® 647 Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab198324)

    • Alexa Fluor® 405 Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab198515)

    • Alexa Fluor® 594 Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab203437)

    • Alexa Fluor® 555 Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab203438)

    • APC Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker (ab225131)

    • PE Anti-M6PR (cation independent) antibody [EPR6599] (ab225132)

    • Anti-M6PR (cation independent) antibody [EPR6599] – BSA and Azide free (ab226090)

  • Compatible Secondaries

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

    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit – Lightning-Link® (ab102918)

    • APC Conjugation Kit – Lightning-Link® (ab201807)

  • Immunizing Peptide (Blocking)

    • M6PR (cation independent) peptide (ab169957)

  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] – Isotype Control (ab172730)

  • Related Products

    • Lysosomal Staining Kit – Blue Fluorescence – Cytopainter (ab112135)

    • Lysosomal Staining Kit – Red Fluorescence – Cytopainter (ab112137)

Applications

The Abpromise guarantee

Our

Abpromise guarantee

covers the use of ab124767 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
WB

(3)

1/50000 – 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa).Can be blocked with

M6PR (cation independent) peptide (ab169957)

.

IHC-P
1/250 – 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.

See

IHC antigen retrieval protocols

.

Flow Cyt
1/100 – 1/500.

ab172730

– Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF

(3)

Use a concentration of 1 µg/ml.
Notes
WB
1/50000 – 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa).Can be blocked with

M6PR (cation independent) peptide (ab169957)

.

IHC-P
1/250 – 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.

See

IHC antigen retrieval protocols

.

Flow Cyt
1/100 – 1/500.

ab172730

– Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
Use a concentration of 1 µg/ml.

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.
  • Xem thêm: Cách tính khoảng cách từ một điểm đến một mặt phẳng-O2 Education

    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

    • Entrez Gene: 25151

      Rat

    • 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

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: M6PR (cation independent) knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: A549 whole cell lysate (20 µg)

    Lanes 1 – 4: Merged signal (red and green). Green – ab124767 observed at 274 kDa. Red – loading control,

    ab18058

    , observed at 130 kDa.

    ab124767 was shown to specifically react with M6PR (cation independent) in wild-type HAP1 cells as signal was lost in M6PR (cation independent) knockout cells. Wild-type and M6PR (cation independent) knockout samples were subjected to SDS-PAGE. Ab124767 and

    ab18058

    (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed

    ab216773

    and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed

    ab216776

    secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab124767 staining M6PR in wild-type HAP1 cells (top panel) and IGF2R knockout HAP1 cells (bottom panel). The cells were fixed with 100% MeOH for 5 min. , permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124767 at 1μg/ml and

    ab195889

     (Mouse monoclonal [DM1A] to alpha Tubulin – Microtubule Marker (Alexa Fluor® 594)) at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (

    ab150081

    ) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/50000 dilution (purified)

    Lane 1 : C6 (rat glioma) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 3 : NIH/3T3 (mouse embryo) whole cell lysate
    Lane 4 : Mouse heart tissue lysate
    Lane 5 : Mouse kidney tissue lysate
    Lane 6 : Mouse spleen tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (

    ab97051

    ) at 1/20000 dilution

    Predicted band size: 274 kDa
    Observed band size: 300 kDa

    why is the actual band size different from the predicted?

    Blocking and diluting buffer 5% NFDM/TBST

  • All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/50000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : Caco-2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 274 kDa

  • All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/200000 dilution (purified)

    Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
    Lane 2 : HEK293 (human embryonic kidney) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (

    ab97051

    ) at 1/100000 dilution

    Predicted band size: 274 kDa
    Observed band size: 300 kDa

    why is the actual band size different from the predicted?

    Blocking and diluting buffer 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded rat colon tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was

    ab97051

    Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

  • Immunohistochemical staining of paraffin embedded mouse colon tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was

    ab97051

    Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

  • Immunohistochemical staining of paraffin embedded human thyroid carcinoma tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was 

    ab97051

    Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

  • Immunohistochemical analysis of formalin fixed, paraffin embedded Human papillary carcinoma tissue section labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/250.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of formalin fixed, paraffin embedded Human tonsil tissue section labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/250.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat (human acute T cell leukemia) cells labelling M6PR with purified ab124767 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100.

    ab150077

    , Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with

    ab7291

    , a mouse anti-tubulin antibody (1/1000) using

    ab150120

    , an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei couterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody was used, followed by anti-mouse secondary antibody (

    ab150120

    ). For negative control 2, mouse primary antibody (

    ab7291

    ) was used followed by anti-rabbit secondary antibody (

    ab150077

    ).  

  • Immunocytochemistry/immunofluorescence analysis of 293T cells labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/100.

  • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labelling M6PR with purified ab124767 at 1/150 (red). Cells were fixed with 4% paraformaldehyde. Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black – Isotype control, rabbit monoclonal IgG. Blue – Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

Protocols

  • Flow cytometry protocols

  • Immunohistochemistry protocols

  • Immunocytochemistry & immunofluorescence protocols

  • Western blot protocols

Click here to view the general protocols

References (41)

ab124767 has been referenced in 41 publications.

  • Curtis ME  et al. Dysregulation of the Retromer Complex System in Down Syndrome. Ann Neurol 88:137-147 (2020).

    PubMed: 32320094

  • Han J  et al. Involvement of CASP9 (caspase 9) in IGF2R/CI-MPR endosomal transport. Autophagy N/A:1-17 (2020).

    PubMed: 32397873

  • Cui L  et al. The Lysosomal Membrane Protein Lamp2 Alleviates Lysosomal Cell Death by Promoting Autophagic Flux in Ischemic Cardiomyocytes. Front Cell Dev Biol 8:31 (2020).

    PubMed: 32117965

  • Grover A  et al. Translational studies of intravenous and intracerebroventricular routes of administration for CNS cellular biodistribution for BMN 250, an enzyme replacement therapy for the treatment of Sanfilippo type B. Drug Deliv Transl Res 10:425-439 (2020).

    PubMed: 31942701

  • Yu XW  et al. A role for CIM6P/IGF2 receptor in memory consolidation and enhancement. Elife 9:N/A (2020).

    PubMed: 32369018

View all Publications for this product

Customer reviews and Q&As

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17 of 7 Abreviews or Q&A

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Western blot abreview for Anti-M6PR (cation independent) antibody [EPR6599]

Excellent

Abreviews

Abreviews

Application
Western blot

Sample
Human Cell lysate – whole cell (HEK293FT cells)

Gel Running Conditions
Reduced Denaturing

Loading amount
10 µg

Specification
HEK293FT cells

Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C

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Abcam user community

Verified customer

Submitted Nov 10 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-M6PR (cation independent) antibody [EPR6599]

Good

Abreviews

Abreviews

Application
Immunocytochemistry/ Immunofluorescence

Sample
Rat Cell (INS1)

Permeabilization
Yes – Tx100

Specification
INS1

Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C

Fixative
Paraformaldehyde

Read More

Abcam user community

Verified customer

Submitted May 21 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-M6PR (cation independent) antibody [EPR6599]

Excellent

Abreviews

Abreviews

Application
Immunocytochemistry/ Immunofluorescence

Sample
Human Cell (HEK 293T cells)

Permeabilization
Yes – 0.1% Saponin, 20% FCS in PBS

Specification
HEK 293T cells

Blocking step
0.1% Saponin, 20% FCS in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 21°C

Fixative
Paraformaldehyde

Read More

Abcam user community

Verified customer

Submitted Jun 18 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-M6PR (cation independent) antibody [EPR6599] – Lysosome Membrane Marker

Excellent

Abreviews

Abreviews

Application
Immunocytochemistry/ Immunofluorescence

Sample
Human Cell (HeLa)

Permeabilization
Yes – Triton X100

Specification
HeLa

Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C

Fixative
Paraformaldehyde

Read More

Abcam user community

Verified customer

Submitted Jun 07 2019

Western blot abreview for Anti-M6PR (cation independent) antibody [EPR6599]

Excellent

Abreviews

Abreviews

Application
Western blot

Sample
Human Cell lysate – other (293T)

Gel Running Conditions
Reduced Denaturing

Loading amount
20 µg

Specification
293T

Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C

Read More

Abcam user community

Verified customer

Submitted Jun 07 2019

Western blot abreview for Anti-Mannose 6 Phosphate Receptor (Cation independent) antibody [EPR6599]

Excellent

Abreviews

Abreviews

Application
Western blot

Sample
Human Cell lysate – whole cell (Hek293)

Loading amount
30 µg

Specification
Hek293

Gel Running Conditions
Reduced Denaturing (10%)

Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Read More

Abcam user community

Verified customer

Submitted Jan 29 2013

Question

Dear All,
a customer of ours is interested in your: Anti-Mannose 6 Phosphate Receptor (Cation independent) antibody [EPR6599] (ab124767) – 100Ul
and she would like to ask if he can proceed by diluting the mother antibody. In case of positive response I would ask if the dilutions should be made in 49% PBS, 0.01% Sodium Azide, 50% Glycerol, 0.05% BSA

Thanks and kind regards

Read More

Answer

Thank you for your enquiry and interest in our products.

We would recommend diluting this antibody for the tested applications. The storage buffer of this antibody contains PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05% so PBS can be used as diluent. Please advise your customer to check the recommended dilution range for the guaranteed applications.

If you need any further assistance in the future, please do not hesitate to contact me.

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