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Product Info Summary
| SKU: | M00949-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsp70 HSPA1A Antibody Picoband® (monoclonal, 3H5)
View all HSP70/HSPA1A Antibodies
SKU/Catalog Number
M00949-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsp70 HSPA1A Antibody Picoband® (monoclonal, 3H5) catalog # M00949-2. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. 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-Hsp70 HSPA1A Antibody Picoband® (monoclonal, 3H5) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00949-2)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
3H5
Isotype
Mouse IgG1
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human Hsp70, different from the related mouse sequence by five amino acids, and from the related rat sequence 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
M00949-2 is reactive to HSPA1A in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
70 kDa
Calculated molecular weight
70.052kDa
Background of HSP70/HSPA1A
HSPA1 (heat shock 70kDa protein 1A) also known as HSP70-1, HSPA1A, HSP70-1A, HSP72 or HSP70I, is a protein that in humans is encoded by the HSPA1A gene. This intronless gene encodes a 70kDa heat shock protein which is a member of the heat shock protein 70 family. The HSPA1A gene encodes a predicted 641-amino acid protein. The HSPA1 gene is mapped on 6p21.33. Shimizu et al. (1999) found that peripheral blood mononuclear cells of 18 major depression patients expressed a short HSPA1A transcript that utilized exon 1 rather than exon 2, which is found in the more common HSPA1A transcript. No protein was associated with expression of this short HSPA1A mRNA, possibly due to lack of a TATA box or loss of internal ribosome binding sites. Treatment with BGP-15, a pharmacologic inducer of Hsp72 that can protect against obesity-induced insulin resistance, improved muscular architecture, strength, and contractile function in severely affected diaphragm muscles in mdx dystrophic mice.
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
M00949-2 is guaranteed for Flow Cytometry, IF, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human Hela whole cell, human COLO-320 whole cell, human SW620 whole cell, human A431 whole cell, human A549 whole cell, human HepG2 whole cell, human PANC-1 whole cell
IHC: human lung cancer tissue
ICC/IF: MCF7 cell
FCM: U20S cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. IHC analysis of Hsp70 using anti-Hsp70 antibody (M00949-2).
Hsp70 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Hsp70 Antibody (M00949-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
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Figure 2. Western blot analysis of Hsp70 using anti-Hsp70 antibody (M00949-2).
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 lysate,
Lane 2: human COLO-320 whole cell lysate,
Lane 3: human SW620 whole cell lysate,
Lane 4: human A431 whole cell lysate,
Lane 5: human A549 whole cell lysate,
Lane 6: human HepG2 whole cell lysate,
Lane 7: human PANC-1 whole cell lysate.
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-Hsp70 antigen affinity purified monoclonal antibody (Catalog # M00949-2) 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.
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Figure 3. Flow Cytometry analysis of U20S cells using anti-Hsp70 antibody (M00949-2).
Overlay histogram showing U20S cells stained with M00949-2 (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-Hsp70 Antibody (M00949-2,1μg/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.
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Figure 4. IF analysis of Hsp70 using anti-Hsp70 antibody (M00949-2).
Hsp70 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-Hsp70 Antibody (M00949-2) 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.
Protein Target Info & Infographic
Gene/Protein Information For HSPA1A (Source: Uniprot.org, NCBI)
Gene Name
HSPA1A
Full Name
Heat shock 70 kDa protein 1A
Weight
70.052kDa
Superfamily
heat shock protein 70 family
Alternative Names
Heat shock 70 kDa protein 1A; Heat shock 70 kDa protein 1B HSPA1A HEL-S-103, HSP70-1, HSP70-1A, HSP70-2, HSP70.1, HSP70.2, HSP70I, HSP72, HSPA1 heat shock protein family A (Hsp70) member 1A heat shock 70 kDa protein 1A|HSP70-1/HSP70-2|HSP70.1/HSP70.2|Heat shock 70 kDa protein 1B|Heat shock 70 kDa protein 2|dnaK-type molecular chaperone HSP70-1|epididymis secretory protein Li 103|epididymis secretory sperm binding protein|heat shock 70 kDa protein 1|heat shock 70 kDa protein 1/2|heat shock 70 kDa protein 1A/1B|heat shock 70kD protein 1A|heat shock 70kDa protein 1A|heat shock-induced protein
*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 HSPA1A, check out the HSPA1A 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 HSPA1A: 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-Hsp70 HSPA1A Antibody Picoband® (monoclonal, 3H5) (M00949-2)
Hello CJ!
M00949-2 has been cited in 5 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Therapeutic efficacy of trehalose eye drops for treatment of murine dry eye induced by an intelligently controlled environmental system
Down-modulation of heat shock protein 70 and up-modulation of Caspase-3 during schisandrin B-induced apoptosis in human hepatoma SMMC-7721 cells
Effects of intracerebral hemorrhage and subsequent minimally invasive hematoma aspiration on expression of apoptosisrelated genes in rats.
Effects of minocycline on the expression of NGF and HSP70 and its neuroprotection role following intracerebral hemorrhage in rats
Yi Sx, Peng Y, Chang Xr, Peng N, Yan J, Lin Yp. World J Gastroenterol. 2007 Apr 21;13(15):2174-8. Effect Of Pre-Moxibustion On Apoptosis And Proliferation Of Gastric Mucosa Cells.
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Customer Q&As
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16 Customer Q&As for Anti-Hsp70 HSPA1A Antibody Picoband® (monoclonal, 3H5)
Question
Thanks for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for uterus using anti-Hsp70 antibody (monoclonal, 3H5) M00949-2. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2020-04-24
Answer
We appreciate 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: 2020-04-24
Question
We are interested in using your anti-Hsp70 antibody (monoclonal, 3H5) for negative regulation of apoptotic process studies. Has this antibody been tested with western blotting on hela whole cell lysate? We would like to see some validation images before ordering.
J. Li
Verified customer
Asked: 2020-04-09
Answer
We appreciate your inquiry. This M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) is tested on human hela, lung cancer tissue, hela whole cell lysate, sw620 whole cell lysate, a431 whole cell lysate, a549 whole cell lysate, hepg2 whole cell lysate, u20s cells. It is guaranteed to work for Flow Cytometry, IHC, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2020-04-09
Question
My question regards to test anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 on rat uterus for research purposes, then I may be interested in using anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2020-04-06
Answer
The products we sell, including anti-Hsp70 antibody (monoclonal, 3H5) M00949-2, 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: 2020-04-06
Question
I have a question about product M00949-2, anti-Hsp70 antibody (monoclonal, 3H5). 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: 2020-01-10
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 M00949-2 anti-Hsp70 antibody (monoclonal, 3H5), 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: 2020-01-10
Question
Would anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 work for WB with uterus?
Verified Customer
Verified customer
Asked: 2019-11-19
Answer
According to the expression profile of uterus, HSPA1A is highly expressed in uterus. So, it is likely that anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 will work for WB with uterus.
Boster Scientific Support
Answered: 2019-11-19
Question
We are currently using anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 for rat tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on monkey tissues as well?
Verified Customer
Verified customer
Asked: 2019-10-30
Answer
The anti-Hsp70 antibody (monoclonal, 3H5) (M00949-2) has not been validated for cross reactivity specifically with monkey 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 monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-10-30
Question
We have seen staining in mouse liver. What should we do? Is anti-Hsp70 antibody (monoclonal, 3H5) supposed to stain liver positively?
Verified Customer
Verified customer
Asked: 2019-10-10
Answer
From literature liver does express HSPA1A. From Uniprot.org, HSPA1A is expressed in endothelial cell, uterus, brain, muscle, pancreas, pns skin, embryonic kidney, brain, cajal-retzius cell fetal brain cortex, cervix carcinoma, cervix carcinoma erythroleukemia, liver, colon carcinoma, among other tissues. Regarding which tissues have HSPA1A expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 23349634
Brain, Muscle, Pancreas, PNS, and Skin, Pubmed ID: 15489334
Cervix carcinoma, Pubmed ID: 17081983, 18691976, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Liver, Pubmed ID: 24275569
Uterus, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-10-10
Question
Our lab used your anti-Hsp70 antibody (monoclonal, 3H5) for Flow Cytometry on liver a few years ago. I am using rat, and We intend to use the antibody for WB next. We want examining liver as well as cervix carcinoma in our next experiment. Do you have any suggestion on which antibody would work the best for WB?
Verified Customer
Verified customer
Asked: 2019-09-23
Answer
I have checked the website and datasheets of our anti-Hsp70 antibody (monoclonal, 3H5) and it appears that M00949-2 has been tested on rat in both Flow Cytometry and WB. Thus M00949-2 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in rat 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: 2019-09-23
Question
My team were content with the WB result of your anti-Hsp70 antibody (monoclonal, 3H5). However we have observed positive staining in embryonic kidney cytoplasm. using this antibody. Is that expected? Could you tell me where is HSPA1A supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-07-16
Answer
From literature, embryonic kidney does express HSPA1A. Generally HSPA1A expresses in cytoplasm. Regarding which tissues have HSPA1A expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 23349634
Brain, Muscle, Pancreas, PNS, and Skin, Pubmed ID: 15489334
Cervix carcinoma, Pubmed ID: 17081983, 18691976, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Liver, Pubmed ID: 24275569
Uterus, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-07-16
Question
I see that the anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2017-11-29
Answer
You can find protocols for WB 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: 2017-11-29
Question
I was wanting to use your anti-Hsp70 antibody (monoclonal, 3H5) for WB for rat uterus 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 rat uterus identification?
A. Johnson
Verified customer
Asked: 2017-05-03
Answer
As indicated on the product datasheet, M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) has been validated for Flow Cytometry, IHC, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat uterus in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-05-03
Question
See below the WB image, lot number and protocol we used for uterus using anti-Hsp70 antibody (monoclonal, 3H5) M00949-2. Please let me know if you require anything else.
R. Edwards
Verified customer
Asked: 2016-11-17
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: 2016-11-17
Question
Is this M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) reactive to the isotypes of HSPA1A?
S. Zhao
Verified customer
Asked: 2016-02-03
Answer
The immunogen of M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) is A synthetic peptide corresponding to a sequence at the C-terminus of human Hsp70 (559-596aa KGKISEADKKKVLDKCQEVISWLDANTLAEKDEFEHKR), different from the related mouse sequence by five amino acids, and from the related rat sequence 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: 2016-02-03
Question
Do you have a BSA free version of anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 available?
A. Banerjee
Verified customer
Asked: 2016-02-02
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-Hsp70 antibody (monoclonal, 3H5) M00949-2 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: 2016-02-02
Question
Does M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
A. Zhang
Verified customer
Asked: 2015-03-06
Answer
You can see on the product datasheet, M00949-2 anti-Hsp70 antibody (monoclonal, 3H5) as been tested on WB. 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: 2015-03-06
Question
Is a blocking peptide available for product anti-Hsp70 antibody (monoclonal, 3H5) (M00949-2)?
O. Krishna
Verified customer
Asked: 2015-02-13
Answer
We do provide the blocking peptide for product anti-Hsp70 antibody (monoclonal, 3H5) (M00949-2). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2015-02-13
