Anti-Hsp70 HSPA1A Antibody Picoband™ (monoclonal, 3H5)

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.

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

Applications

M00949-2 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

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.

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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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry, 1-3μg/1x10⁶ cells

Validation Images & Assay Conditions

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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).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.

Gene/Protein Information For HSPA1A (Source: Uniprot.org, NCBI)

Alternative Names

dnaK-type molecular chaperone HSP70-1; FLJ54303; FLJ54370; FLJ54392; FLJ54408; FLJ75127; 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; HSP70; HSP70.1/HSP70.2; HSP70-1; HSP70-1/HSP70-2; HSP70-1A; HSP70I; HSP72; HSPA1; HSPA1A; HSPA1B 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

For more info on HSPA1A, check out the HSPA1A Infographic

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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].

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