Product Info Summary
SKU: | A00241-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Histone H2A.X/H2AFX Antibody Picoband™
SKU/Catalog Number
A00241-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Histone H2A.X/H2AFX Antibody Picoband™ catalog # A00241-1. Tested in ELISA, Flow Cytometry, IHC, 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-Histone H2A.X/H2AFX Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00241-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Histone H2A.X/H2AFX recombinant protein (Position: S2-T121).
*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
A00241-1 is reactive to H2AX in Human, Mouse, Rat
Applications
A00241-1 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Observed Molecular Weight
15 kDa
Calculated molecular weight
15.145kDa
Background of H2AX
H2A histone family member X (usually abbreviated asH2AX) is a type of histone protein from the H2A family encoded by the H2AFX gene. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a replication-independent histone that is a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif.
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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Flow Cytometry, 1-3μg/1x106 cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of H2AFX using anti-H2AFX antibody (A00241-1).
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 HepG2 whole cell lysates
Lane 2: human Caco-2 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 rabbit anti-H2AFX antigen affinity purified polyclonal antibody (Catalog # A00241-1) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for H2AFX at approximately 15KD. The expected band size for H2AFX is at 15KD.
Click image to see more details
Figure 2. IHC analysis of H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of human lung cancer tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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 H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of rat small intestine tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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 H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of human mammary cancer tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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. IHC analysis of H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of human rectal cancer tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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 6. IHC analysis of H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of mouse brain tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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 7. IHC analysis of H2AFX using anti-H2AFX antibody (A00241-1).
H2AFX was detected in paraffin-embedded section of mouse small intestine tissues. 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 1μg/ml rabbit anti-H2AFX Antibody (A00241-1) 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 8. Flow Cytometry analysis of HepG2 cells using anti-H2AFX antibody (A00241-1).
Overlay histogram showing HepG2 cells stained with A00241-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-H2AFX Antibody (A00241-1,1μg/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 9. Western blot analysis of H2AFX using anti-H2AFX antibody (A00241-1).
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 Caco-2 whole cell lysates,
Lane 2: human HepG2 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 rabbit anti-H2AFX antigen affinity purified polyclonal antibody (Catalog # A00241-1) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for H2AFX at approximately 15KD. The expected band size for H2AFX is at 15KD.
Protein Target Info & Infographic
Gene/Protein Information For H2AX (Source: Uniprot.org, NCBI)
Gene Name
H2AX
Full Name
Histone H2AX
Weight
15.145kDa
Superfamily
histone H2A family
Alternative Names
H2A histone family, member X; H2A.X; H2a/x; H2AX histone; H2AXhistone H2A.x; YH2AX H2AX H2A.X, H2A/X, H2AFX H2A.X variant histone histone H2AX|H2A histone family member X|H2AX histone|histone H2A.x
*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 H2AX, check out the H2AX 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 H2AX: 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-Histone H2A.X/H2AFX Antibody Picoband™ (A00241-1)
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No publications found for A00241-1
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Customer Reviews
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1 Reviews For Anti-Histone H2A.X/H2AFX Antibody Picoband™
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Immunohistochemistry of Anti-Histone H2A.X/H2AFX Antibody
Excellent
Application | Immunohistochemistry (paraffin-embedded ) |
---|---|
Blocking step | 5% BSA as a blocking agent for 30 min at 37°C |
Sample | Human colon |
Fixative | Fixed with 4% paraformaldehyde |
Primary Ab Incubation | 37 °C for 30 minutes |
Primary Ab Incubation diluent | 5% BSA in TBS |
Primary Ab Concentration | 2ug/ml |
Secondary Antibody | SABC
kit from Boster Bio, (SA1022 |
Secondary Ab Dilution | The kit was ready to use, no dilution needed |
Secondary Ab Incubation | at 37°C for 30 min |
Verified Customer
Verified customer
Submitted 2019-02-21
Customer Q&As
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1 Customer Q&As for Anti-Histone H2A.X/H2AFX Antibody Picoband™
Question
We are currently using anti-Histone H2A.X/H2AFX antibody A00241-1 for human tissue, and we are happy with the ELISA results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on monkey tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-09
Answer
The anti-Histone H2A.X/H2AFX antibody (A00241-1) has not been tested for cross reactivity specifically with monkey tissues, but 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: 2019-08-09