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Product Info Summary
| SKU: | M01891-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Rab5/RAB5A Antibody Picoband® (monoclonal, 3E9)
SKU/Catalog Number
M01891-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Rab5/RAB5A Antibody Picoband® (monoclonal, 3E9) catalog # M01891-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Rab5/RAB5A Antibody Picoband® (monoclonal, 3E9) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01891-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
Clone: 3E9
Isotype
Mouse IgG2b
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human Rab5, different from the related mouse and rat sequences by three amino acids.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01891-1 is reactive to RAB5A in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
24 kDa
Calculated molecular weight
25035 MW
Background of RAB5A
RAB5A (Ras-associated protein RAB5A), also called RAB5, is a protein that in humans is encoded by the RAB5A gene. RAB5 is a rate-limiting component of the machinery regulating the kinetics of membrane traffic in the early endocytic pathway. The RAB5A gene is mapped on 3p24.3. RAB5 is indispensable for a form of receptor tyrosine kinase-induced actin remodeling called circular ruffling.It signals to the actin cytoskeleton through RNTRE, a RAB5-specific GTPase-activating protein (GAP).RAB5 activity on phagosome membranes began to increase on disassembly of the actin coat encapsulating phagosomes. In addition, RAB5 activation is either continuous or repetitive for up to 10 minutes, but it ends before the collapse of engulfed apoptotic cells. Expression of a dominant-negative mutant of RAB5 delayed this collapse of apoptotic thymocytes, showing a role for RAB5 in phagosome maturation.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M01891-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human Hela whole cell, human placenta tissue, human A549 whole cellhuman A375 whole cell, human SW620 whole cell, human U20S whole cell, human HEK293 whole cell, human U87 whole cell, rat brain tissue, mouse brain tissue, mouse Neuro-2a whole cell
ICC/IF: MCF7 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. IF analysis of RAB5A using anti-RAB5A antibody (M01891-1).
RAB5A was detected in immunocytochemical section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-RAB5A Antibody (M01891-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 2. Western blot analysis of Rab5/RAB5A using anti-Rab5/RAB5A antibody (M01891-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human A549 whole cell lysates.
Lane 4: human A375 whole cell lysates,
Lane 5: human SW620 whole cell lysates,
Lane 6: human U20S whole cell lysates,
Lane 7: human HEK293 whole cell lysates,
Lane 8: human U87 whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: mouse brain tissue lysates,
Lane 11: mouse Neuro-2a whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Rab5/RAB5A antigen affinity purified monoclonal antibody (Catalog # M01891-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Rab5/RAB5A at approximately 24KD. The expected band size for Rab5/RAB5A is at 24KD.
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Figure 3. Flow Cytometry analysis of A431 cells using anti-Rab5/RAB5A antibody (M01891-1).
Overlay histogram showing A431 cells stained with M01891-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Rab5/RAB5A Antibody (M01891-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For RAB5A (Source: Uniprot.org, NCBI)
Gene Name
RAB5A
Full Name
Ras-related protein Rab-5A
Weight
25035 MW
Superfamily
small GTPase superfamily
Alternative Names
Ras-related protein Rab-5A; RAB5A; RAB5 RAB5A RAB5 RAB5A, member RAS oncogene family ras-related protein Rab-5A|RAS-associated protein RAB5A
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on RAB5A, check out the RAB5A Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for RAB5A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Rab5/RAB5A Antibody Picoband® (monoclonal, 3E9) (M01891-1)
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Customer Q&As
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15 Customer Q&As for Anti-Rab5/RAB5A Antibody Picoband® (monoclonal, 3E9)
Question
Can you help my question with product M01891-1, anti-Rab5/RAB5A antibody (monoclonal, 3E9). I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2020-04-16
Answer
It is not recommended storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9), we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2020-04-16
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for substantia nigra using anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-12-19
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-12-19
Question
Would M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9) work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-10-04
Answer
As indicated on the product datasheet, M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9) as been tested on Flow Cytometry. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2019-10-04
Question
We have seen staining in human substantia nigra. What should we do? Is anti-Rab5/RAB5A antibody (monoclonal, 3E9) supposed to stain substantia nigra positively?
Verified Customer
Verified customer
Asked: 2019-09-30
Answer
From literature substantia nigra does express RAB5A. From Uniprot.org, RAB5A is expressed in substantia nigra, brain, brain subthalamic nucleus, cervix placenta, cervix carcinoma, melanoma, among other tissues. Regarding which tissues have RAB5A expression, here are a few articles citing expression in various tissues:
Brain, and Subthalamic nucleus, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 11062261, 18669648
Cervix, and Placenta, Pubmed ID: 15489334
Melanoma, Pubmed ID: 17081065
Boster Scientific Support
Answered: 2019-09-30
Question
We are currently using anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 for rat tissue, and we are satisfied with the Flow Cytometry results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on feline tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-08
Answer
The anti-Rab5/RAB5A antibody (monoclonal, 3E9) (M01891-1) has not been tested for cross reactivity specifically with feline tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-08-08
Question
I see that the anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 works with Flow Cytometry, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-05
Answer
You can find protocols for Flow Cytometry on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-06-05
Question
Would anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 work for Flow Cytometry with substantia nigra?
Verified Customer
Verified customer
Asked: 2019-04-16
Answer
According to the expression profile of substantia nigra, RAB5A is highly expressed in substantia nigra. So, it is likely that anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 will work for Flow Cytometry with substantia nigra.
Boster Scientific Support
Answered: 2019-04-16
Question
My colleagues were satisfied with the WB result of your anti-Rab5/RAB5A antibody (monoclonal, 3E9). However we have observed positive staining in substantia nigra cell membrane using this antibody. Is that expected? Could you tell me where is RAB5A supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-11-08
Answer
From what I have seen in literature, substantia nigra does express RAB5A. Generally RAB5A expresses in cell membrane. Regarding which tissues have RAB5A expression, here are a few articles citing expression in various tissues:
Brain, and Subthalamic nucleus, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 11062261, 18669648
Cervix, and Placenta, Pubmed ID: 15489334
Melanoma, Pubmed ID: 17081065
Boster Scientific Support
Answered: 2018-11-08
Question
I was wanting to use your anti-Rab5/RAB5A antibody (monoclonal, 3E9) for Flow Cytometry for human substantia nigra on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for human substantia nigra identification?
K. Zhao
Verified customer
Asked: 2018-01-17
Answer
As indicated on the product datasheet, M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9) has been validated for Flow Cytometry, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human substantia nigra in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-01-17
Question
Is a blocking peptide available for product anti-Rab5/RAB5A antibody (monoclonal, 3E9) (M01891-1)?
M. Johnson
Verified customer
Asked: 2017-11-23
Answer
We do provide the blocking peptide for product anti-Rab5/RAB5A antibody (monoclonal, 3E9) (M01891-1). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2017-11-23
Question
I am interested in to test anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 on human substantia nigra for research purposes, then I may be interested in using anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
E. Brown
Verified customer
Asked: 2017-02-10
Answer
The products we sell, including anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2017-02-10
Question
Is there a BSA free version of anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 available?
D. Lewis
Verified customer
Asked: 2016-07-12
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2016-07-12
Question
Is this M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9) reactive to the isotypes of RAB5A?
R. Evans
Verified customer
Asked: 2015-06-15
Answer
The immunogen of M01891-1 anti-Rab5/RAB5A antibody (monoclonal, 3E9) is A synthetic peptide corresponding to a sequence at the C-terminus of human Rab5 (179-215aa KKLPKNEPQNPGANSARGRGVDLTEPTQPTRNQCCSN), different from the related mouse and rat sequences by three amino acids. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2015-06-15
Question
Our lab used your anti-Rab5/RAB5A antibody (monoclonal, 3E9) for Flow Cytometry on cervix carcinoma last year. I am using human, and I plan to use the antibody for WB next. My lab would like examining cervix carcinoma as well as substantia nigra in our next experiment. Do you have any suggestion on which antibody would work the best for WB?
K. Moore
Verified customer
Asked: 2015-03-04
Answer
I looked at the website and datasheets of our anti-Rab5/RAB5A antibody (monoclonal, 3E9) and it appears that M01891-1 has been tested on human in both Flow Cytometry and WB. Thus M01891-1 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2015-03-04
Question
Please see the WB image, lot number and protocol we used for substantia nigra using anti-Rab5/RAB5A antibody (monoclonal, 3E9) M01891-1. Please let me know if you require anything else.
E. Wu
Verified customer
Asked: 2014-03-20
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2014-03-20
