Product Info Summary
SKU: | PB9641 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsc70/HSPA8 Antibody Picoband™
View all HSPA8/HSC71/Hsc70 Antibodies
SKU/Catalog Number
PB9641
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsc70/HSPA8 Antibody Picoband™ catalog # PB9641. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
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-Hsc70/HSPA8 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9641)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Hsc70 recombinant protein (Position: Q520-A614). Human Hsc70 shares 98.9% amino acid (aa) sequence identity with both mouse and rat Hsc70.
*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
PB9641 is reactive to HSPA8 in Human, Mouse, Rat
Applications
PB9641 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
71 kDa
Calculated molecular weight
70.898kDa
Background of HSPA8/HSC71/Hsc70
HSPA8 (heat shock 70kDa protein 8) also known as HSC70, HSC71, HSP73, HSPA10, FORMERLY, LAP1 or LPS-ASSOCIATED PROTEIN 1, is a heat shock protein that in humans is encoded by the HSPA8 gene. The HSPA8 gene contains 9 exons and spans 5 kb. The deduced HSPA8 protein has 646 amino acids and a predicted molecular mass of 70,899 Da. And the HSPA8 gene is mapped on 11q24.1. HSPA8 plays an important role in cells by transiently associating with nascent polypeptides to facilitate correct folding. HSP73 also functions as an ATPase in the disassembly of clathrin-coated vesicles during transport of membrane components through the cell. Rapid decay involves AU-rich binding protein AUF1, which complexes with heat-shock proteins HSC70 and HSP70, translation initiation factor EIF4G, and poly (A)-binding protein. In the absence of Il3, Hsc70 formed a complex with Hsp40 and Hip, and this complex, in association with Eif4g and Pabp, formed a high-stability complex with Bim mRNA that protected it from ribonucleases.
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
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, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human CACO-2 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human THP-1 whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: human PC-3 whole cell lysates,
Lane 8: human U87MG whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hsc70 antigen affinity purified polyclonal antibody (Catalog # PB9641) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsc70 at approximately 71 kDa. The expected band size for Hsc70 is at 71 kDa.
Click image to see more details
Figure 2. IHC analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
Hsc70 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsc70 Antibody (PB9641) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
Hsc70 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsc70 Antibody (PB9641) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
Hsc70 was detected in a paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsc70 Antibody (PB9641) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IF analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
Hsc70 was detected in immunocytochemical section of U20S 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 rabbit anti-Hsc70 Antibody (PB9641) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
Click image to see more details
Figure 6. Flow Cytometry analysis of K562 cells using anti-Hsc70 antibody (PB9641).
Overlay histogram showing K562 cells stained with PB9641 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Hsc70 Antibody (PB9641,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 7. Western blot analysis of Hsc70 using anti-Hsc70 antibody (PB9641).
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 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: rat spleen tissue lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse spleen tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hsc70 antigen affinity purified polyclonal antibody (Catalog # PB9641) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsc70 at approximately 71 kDa. The expected band size for Hsc70 is at 71 kDa.
Protein Target Info & Infographic
Gene/Protein Information For HSPA8 (Source: Uniprot.org, NCBI)
Gene Name
HSPA8
Full Name
Heat shock cognate 71 kDa protein
Weight
70.898kDa
Superfamily
heat shock protein 70 family
Alternative Names
Heat shock 70 kDa protein 8; heat shock 70kD protein 8; heat shock 70kDa protein 8; heat shock cognate 71 kDa protein; heat shock cognate protein 54; heat shock cognate protein, 71-kDa; HSC54; HSC70; HSC70heat shock 70kd protein 10; HSC71; HSP71; HSP73; HSP73constitutive heat shock protein 70; HSPA10; HSPA8; LAP1; lipopolysaccharide-associated protein 1; LPS-associated protein 1; MGC131511; MGC29929; NIP71; N-myristoyltransferase inhibitor protein 71 HSPA8 HEL-33, HEL-S-72p, HSC54, HSC70, HSC71, HSP71, HSP73, HSPA10, LAP-1, LAP1, NIP71 heat shock protein family A (Hsp70) member 8 heat shock cognate 71 kDa protein|LPS-associated protein 1|N-myristoyltransferase inhibitor protein 71|constitutive heat shock protein 70|epididymis luminal protein 33|epididymis secretory sperm binding protein Li 72p|heat shock 70kDa protein 8|heat shock 70kd protein 10|heat shock cognate protein 54|lipopolysaccharide-associated protein 1
*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 HSPA8, check out the HSPA8 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 HSPA8: 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-Hsc70/HSPA8 Antibody Picoband™ (PB9641)
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Customer Q&As
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16 Customer Q&As for Anti-Hsc70/HSPA8 Antibody Picoband™
Question
Is there a BSA free version of anti-Hsc70/HSPA8 antibody PB9641 available?
Verified Customer
Verified customer
Asked: 2020-04-29
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Hsc70/HSPA8 antibody PB9641 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: 2020-04-29
Question
I was wanting to use your anti-Hsc70/HSPA8 antibody for WB for rat colon carcinoma ovarian carcinoma on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for rat colon carcinoma ovarian carcinoma identification?
S. Carter
Verified customer
Asked: 2020-04-22
Answer
As indicated on the product datasheet, PB9641 anti-Hsc70/HSPA8 antibody has been validated for IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat colon carcinoma ovarian carcinoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-04-22
Question
I have a question about product PB9641, anti-Hsc70/HSPA8 antibody. 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: 2020-03-13
Answer
We do not advise 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 PB9641 anti-Hsc70/HSPA8 antibody, 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: 2020-03-13
Question
Here is the WB image, lot number and protocol we used for colon carcinoma ovarian carcinoma using anti-Hsc70/HSPA8 antibody PB9641. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2020-03-11
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: 2020-03-11
Question
I see that the anti-Hsc70/HSPA8 antibody PB9641 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2020-02-24
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: 2020-02-24
Question
We have observed staining in mouse brain. Are there any suggestions? Is anti-Hsc70/HSPA8 antibody supposed to stain brain positively?
Verified Customer
Verified customer
Asked: 2019-12-05
Answer
Based on literature brain does express HSPA8. Based on Uniprot.org, HSPA8 is expressed in frontal cortex, placenta skin, embryonic kidney, colon carcinoma ovarian carcinoma, brain, cajal-retzius cell fetal brain cortex, lymphoblast, melanoma, leukemic t-cell, cervix carcinoma, cervix carcinoma erythroleukemia, liver, colon carcinoma, among other tissues. Regarding which tissues have HSPA8 expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 1286667
Cervix carcinoma, Pubmed ID: 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Embryonic kidney, Pubmed ID: 17525332
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lymphoblast, Pubmed ID: 14654843
Melanoma, Pubmed ID: 17081065
Placenta, and Skin, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2019-12-05
Question
Will anti-Hsc70/HSPA8 antibody PB9641 work for WB with colon carcinoma ovarian carcinoma?
Verified Customer
Verified customer
Asked: 2019-06-03
Answer
According to the expression profile of colon carcinoma ovarian carcinoma, HSPA8 is highly expressed in colon carcinoma ovarian carcinoma. So, it is likely that anti-Hsc70/HSPA8 antibody PB9641 will work for WB with colon carcinoma ovarian carcinoma.
Boster Scientific Support
Answered: 2019-06-03
Question
Will PB9641 anti-Hsc70/HSPA8 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-05-24
Answer
As indicated on the product datasheet, PB9641 anti-Hsc70/HSPA8 antibody 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: 2019-05-24
Question
I would like using your anti-Hsc70/HSPA8 antibody for regulation of protein import studies. Has this antibody been tested with western blotting on mouse thymus tissue? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-11-14
Answer
Thanks for your inquiry. This PB9641 anti-Hsc70/HSPA8 antibody is validated on rat thymus tissue, liver tissue, tissue lysate, testis tissue, mouse thymus tissue, hela whole cell lysate, 293t whole cell lysate. It is guaranteed to work for IHC, 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: 2018-11-14
Question
We are currently using anti-Hsc70/HSPA8 antibody PB9641 for human tissue, and we are satisfied 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: 2018-05-18
Answer
The anti-Hsc70/HSPA8 antibody (PB9641) has not been tested 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: 2018-05-18
Question
We need to test anti-Hsc70/HSPA8 antibody PB9641 on rat colon carcinoma ovarian carcinoma for research purposes, then I may be interested in using anti-Hsc70/HSPA8 antibody PB9641 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
V. Parker
Verified customer
Asked: 2017-12-14
Answer
The products we sell, including anti-Hsc70/HSPA8 antibody PB9641, 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: 2017-12-14
Question
Is a blocking peptide available for product anti-Hsc70/HSPA8 antibody (PB9641)?
Verified Customer
Verified customer
Asked: 2017-10-20
Answer
We do provide the blocking peptide for product anti-Hsc70/HSPA8 antibody (PB9641). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2017-10-20
Question
Is this PB9641 anti-Hsc70/HSPA8 antibody reactive to the isotypes of HSPA8?
Verified Customer
Verified customer
Asked: 2017-09-21
Answer
The immunogen of PB9641 anti-Hsc70/HSPA8 antibody is E.coli-derived human Hsc70 recombinant protein (Position: Q520-A614). Human Hsc70 shares 98.9% amino acid (aa) sequence identity with both mouse and rat Hsc70. 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: 2017-09-21
Question
We ordered your anti-Hsc70/HSPA8 antibody for IHC on lymphoblast in the past. I am using rat, and I plan to use the antibody for WB next. We are interested in examining lymphoblast as well as frontal cortex in our next experiment. Could you please give me some suggestion on which antibody would work the best for WB?
C. Dhar
Verified customer
Asked: 2015-08-21
Answer
I have checked the website and datasheets of our anti-Hsc70/HSPA8 antibody and it appears that PB9641 has been validated on rat in both IHC and WB. Thus PB9641 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: 2015-08-21
Question
Thanks for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for colon carcinoma ovarian carcinoma using anti-Hsc70/HSPA8 antibody PB9641. Let me know if you need anything else.
R. Parker
Verified customer
Asked: 2015-06-12
Answer
Thanks 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: 2015-06-12
Question
My team were satisfied with the WB result of your anti-Hsc70/HSPA8 antibody. However we have seen positive staining in cervix carcinoma nucleolus. using this antibody. Is that expected? Could you tell me where is HSPA8 supposed to be expressed?
K. Dhar
Verified customer
Asked: 2014-07-03
Answer
Based on literature, cervix carcinoma does express HSPA8. Generally HSPA8 expresses in cytoplasm. melanosome. nucleus, nucleolus. Regarding which tissues have HSPA8 expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 1286667
Cervix carcinoma, Pubmed ID: 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Embryonic kidney, Pubmed ID: 17525332
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lymphoblast, Pubmed ID: 14654843
Melanoma, Pubmed ID: 17081065
Placenta, and Skin, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2014-07-03