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H1.5 Linker Histone, Cluster Member Peptide 1

Antibody: CPTC-H1-5-1
Background

Catalog Number:

CPTC-H1-5-1

Target Antigen:

H1.5 Linker Histone, Cluster Member Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

12/06/2024

Antigen Recognition(s):

Peptide, Endogenous

Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image Affinity and binding kinetics of CPTC-H1-5-1 and BSA conjugated peptide, CAPVEK-pS-PAK, were measured using biolayer interferometry. BSA-conjugated peptide was amine coupled to the AR2G biosensors. The CPTC-H1-5-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. All binding data were analyzed globally using a 1:2 (bivalent) fitting model. Click image to enlarge

CPTC-H1-5-1 Affinity and Kinetics (Bio-Layer Interferometry) 

Result: High Binding

Affinity and binding kinetics of CPTC-H1-5-1 and BSA conjugated peptide, CAPVEK-pS-PAK, were measured using biolayer interferometry. BSA-conjugated peptide was amine coupled to the AR2G biosensors. The CPTC-H1-5-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without peptide were used as references for background subtraction. All binding data were analyzed globally using a 1:2 (bivalent) fitting model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-H1-5-1 and BSA-conjugated peptide, CAPVEK-pS-PAK, were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-H1-5-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. All data were double referenced and analyzed globally using a 1:2 (bivalent) fitting model. Click image to enlarge

CPTC-H1-5-1 Affinity and Kinetics (Surface Plasmon Resonance)

Result: High Binding

Affinity and binding kinetics of CPTC-H1-5-1 and BSA-conjugated peptide, CAPVEK-pS-PAK, were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled onto a Series S CM5 biosensor chip. CPTC-H1-5-1 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. All data were double referenced and analyzed globally using a 1:2 (bivalent) fitting model.


Characterization SOP Files

  Flow Cytometry

CPTC-H1-5-1 (Flow Cytometry)

Result: Negative

Flow cytometric analysis of thyroid medullarry TT cells, asynchronous (blue) or synchronous (serum starved 24 hours, green) using CPTC-H1-5-1 rabbit antibody (solid lines) or concentration matched rabbit isotype control antibody (dashed lines). TT cells were permeabilized, fixed, and then stained with either CPTC-H1-5-1 or rabbit isotype control antibody. A BV421 conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo.


Characterization SOP Files

  IHC Tissue

CPTC-H1-5-1 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-H1-5-1 as primary antibody against BSA-conjugated peptide, CAPVEK-pS-PAK, coated on the plate and detected using the goat anti-rabbit antibody and TMB. Click image to enlarge

CPTC-H1-5-1 Indirect ELISA

Result: High Binding

Indirect ELISA using CPTC-H1-5-1 as primary antibody against BSA-conjugated peptide, CAPVEK-pS-PAK, coated on the plate and detected using the goat anti-rabbit antibody and TMB.


Characterization SOP Files

  NCI 60 Protein Array
Click to enlarge image Protein Array in which CPTC-H1-5-1 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red). Click image to enlarge

CPTC-H1-5-1 NCI 60 Protein Array

Result: Positive

Protein Array in which CPTC-H1-5-1 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red).


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate

CPTC-H1-5-1 WB (Over-expressed lysates)

Result: Negative

Western Blot using CPTC-H1-5-1 as primary antibody against the over-expressed lysate of H1-5. The antibody is not able to recognize the target protein in the tested lysate.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-H1-5-1 WB (Cell lysates)

Result: Negative

Western blot using CPTC-H1-5-1 as primary antibody against the cell lysates of TT, H146 and H1184 cells, asynchronous (+) or synchronous - serum starved 24hours (-). The antibody was not able to detect the target protein in any of the tested cell lysates. The same membrane was probed with an anti CytC antibody, and all the cell lysates show the presence of the housekeeping protein.


Characterization SOP Files

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Antigen: H1.5 Linker Histone, Cluster Member Peptide 1
Background

NCI Identification Number:

00536

Antigen Name:

H1.5 Linker Histone, Cluster Member Peptide 1

CPTC Name:

CPTC-H1-5 Peptide 1

Aliases:

H1.5 Linker Histone, Cluster Member; HIST1H1B; H1s-3; H1F5; Histone Cluster 1 H1 Family Member B; H1 Histone Family, Member 5; Histone Cluster 1, H1b; Histone 1, H1b; Histone H1s-3; Histone H1.5; Histone H1a; Histone H1b; H1b; H1.5; H1B; H1

Function:

Histones are basic nuclear proteins responsible for 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 is intronless and encodes a replication-dependent histone that is a member of the histone H1 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the small histone gene cluster on chromosome 6p22-p21.3. H1-5 (H1.5 Linker Histone, Cluster Member) is a Protein Coding gene. Diseases associated with H1-5 include Retinitis Pigmentosa 18 and Tonsil Squamous Cell Carcinoma. Among its related pathways are Cellular responses to stimuli and Programmed Cell Death. An important paralog of this gene is H1-4.Histone H1 protein binds to linker DNA between nucleosomes forming the macromolecular structure known as the chromatin fiber. Histones H1 are necessary for the condensation of nucleosome chains into higher-order structured fibers. Acts also as a regulator of individual gene transcription through chromatin remodeling, nucleosome spacing and DNA methylation (By similarity). ( H15_HUMAN,P16401 )

Chromosomal Localization:

6p22.1

Accession Number:

UniProt Accession Number:

P16401

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

CAPVEK-pS-PAK

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1100

Last Updated:

11/18/2022

Links

Characterization Data

SOPs

No SOPs available.

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