Human SSBP1 Antibody

R&D Systems | Catalog # AF6588

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all SSBP1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human SSBP1
Met1-Glu148
Accession # Q04837

Specificity

Detects human SSBP1 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human SSBP1 Antibody

Detection of Human SSBP1 antibody by Western Blot.

Detection of Human SSBP1 by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line, MOLT-4 human acute lymphoblastic leukemia cell line, K562 human chronic myelogenous leukemia cell line, and UT-7 human acute myeloid leukemia cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human SSBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6588) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for SSBP1 at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
SSBP1 antibody in HL-60 Human Cell Line by Immunocytochemistry (ICC).

SSBP1 in HL‑60 Human Cell Line.

SSBP1 was detected in immersion fixed HL-60 human acute promyelocytic leukemia cell line using Sheep Anti-Human SSBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6588) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of SSBP1 by Western Blot

Detection of SSBP1 by Western Blot

Outside–in degradation of mitochondrial proteins.A, mitochondrial proteins were enriched among significantly regulated proteins. B, abundance profiles of significantly regulated proteins from various mitochondrial subcompartments show trends dependent on parkin activity. Indicated are size of total annotated proteins (left; dark profiles) and those after filtering (right; bright profiles); four mitochondrial proteins are of unknown or dual sublocalization (data not shown). C, average profile of each cluster sorted by half maximum in combination with overrepresentation analysis of mitochondrial subcompartments allows for the discrimination of “early” degraded MOM, “intermediate” degraded MOM/IMS/MIM, and “late” degraded MIM/matrix proteins. D, Western blot detection of mitochondrial subcompartment proteins during mitophagy validates stepwise degradation of mitochondrial subcompartments. IMS, inter membrane space; Mito, mitochondrion; MIM, mitochondrial inner membrane; MOM, mitochondrial outer membrane. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S1535947621001638), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SSBP1 by Western Blot

Detection of SSBP1 by Western Blot

Proteasome action is required at late stages of mitophagy. (A) Scheme of treatment time points for proteasome inhibition experiments. Bold text indicates treatment with CCCP while normal text indicates treatment with proteasome inhibitor MG132. (B and C) Western blot analysis of proteasome influence on the indicated inner mitochondrial proteins. (D) Representative images of WT-parkin HeLa cells showing SSBP1 (magenta) and TOM20 (green) after 24 h of CCCP treatment or CCCP treatment combined with MG132 added after 8 h of mitochondria depolarization. Scale bar = 5 µm. (E and F) Single cell analysis of (E) mean number of SSSP1 clumps and (F) relative average SSBP1 clump area per cell. Bars: Mean ± SEM. ns: non-significant; *: p≤0.05, ***: p≤0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40396016), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SSBP1 by Immunohistochemistry

Detection of SSBP1 by Immunohistochemistry

Parkin activity is essential for the removal of depolarized mitochondria.A, immunofluorescence staining of WT-parkin and C431A-parkin cells after early (2 h) and late (18 h) depolarization times. Scale bars represent 5 μm. B, experimental setup combining subcellular protein fractionation with peptide dimethyl labeling and high pH reversed-phase fractionation during “early” and “late” mitophagy stages in WT-parkin and C431A-parkin cells. Basic workflow applied for proteome and phosphoproteome analysis. C, correlation plot between replicates after log2 transformation of protein ratios. Common light dimethyl labeling channel (0 h postdepolarization) allows comparison between “early” and “late” triple dimethyl-labeling sets per cell line. Parkin-dependent degradation of mitochondrial proteins reflected by shifting data points (density colored points) from “unregulated” toward “downregulated” in WT-parkin expressing cells in comparison to total proteome (gray points). Number of valid pairs per condition and Spearman rank correlation (in brackets) indicated for total (T) and mitochondrial annotated proteins (M). Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S1535947621001638), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SSBP1 by Immunohistochemistry

Detection of SSBP1 by Immunohistochemistry

Autophagy activation and delivery to lysosomes of inner- mitochondrial material. (A and B) Influence of BafA inhibition on TOM20-negative mitochondrial-aggresomes. (A) Representative immunofluorescence images showing TOM20 (green) and SSBP1 (magenta) structures at the indicated CCCP-treatment time points in combination with autophagy inhibition (BafA). (B) Single-cell analysis of mean number of mitochondrial SSBP1 and TOM20 puncta in (A). (C) COX IV-positive puncta (magenta) are positive for EGFP-LC3B (yellow). Arrows indicate COX IV puncta partially engaged with EGFP-LC3B structures. Closed arrowheads indicate complete overlap of COX IV puncta and EGFP-LC3B. Open arrowheads indicate non co-localizing COX IV puncta with EGFP-LC3B. (D) Representative immunofluorescence images showing COX IV-positive puncta (green) co-localizing with LysoTracker (magenta) at the indicated time points. Arrows indicate mitochondrial puncta while arrowheads indicate inner mitochondrial aggregates. (E and F) Representative immunofluorescence images showing (E) COX IV-positive (magenta) or (F) SSBP1-positive mitochondrial aggregates (magenta) close to LAMP1 patches (green) at the indicated time points. Arrows indicate mitochondrial puncta while arrowheads indicate inner mitochondrial aggregates. ****: p ≤0.0001. All scale bars = 5 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40396016), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human SSBP1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HL-60 human acute promyelocytic leukemia cell line

Western Blot

1 µg/mL
Sample: HEK293 human embryonic kidney cell line, MOLT‑4 human acute lymphoblastic leukemia cell line, K562 human chronic myelogenous leukemia cell line, and UT‑7 human acute myeloid leukemia cell line

Reviewed Applications

Read 1 review rated 4 using AF6588 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: SSBP1

SSBP1 (Single-Stranded DNA –Binding Protein; also MtSSB and PWP1-interacting protein 17) is a 17-18 kDa member of the mtSSB protein family. SSBP1 should not be confused with SSB1/SOSS-B1. Although SSBP1 binds single-stranded mitochondrial DNA, it is not the product of a mitochondrial gene. Rather, it is synthesized on ribosomes in the cytosol, and undergoes transport into mitochondria via an N-terminal mitochondrial localization signal. After import, the localization motif is cleaved, allowing for homotetramerization. Upon DNA replication, SSBP1 is phosphorylated by CDK, allowing for DNA duplication. Human SSBP1 is 148 amino acids (aa) in length. It contains a cleavable mitochondrial targeting sequence (aa 1-16), an SSB domain (aa 30-141), one phosphorylation (Ser79) and one acetylation site (Lys113). There is one potential isoform that shows a 25 aa substitution for aa 135-148. Full-length human SSBP1 shares 92% aa identity with mouse SSBP1.

Long Name

Single-stranded DNA Binding Protein Motochondrial

Alternate Names

mitochondrial, MtSSB, single-stranded DNA binding protein 1

Entrez Gene IDs

6742 (Human); 381760 (Mouse); 54304 (Rat)

Gene Symbol

SSBP1

UniProt

Q04837

Additional SSBP1 Products

Product Documents for Human SSBP1 Antibody

Certificate of Analysis

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Product Specific Notices for Human SSBP1 Antibody

For research use only

Citations for Human SSBP1 Antibody

Customer Reviews for Human SSBP1 Antibody (1)

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  • Human SSBP1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: H1299 human lung carcinoma cell line, H1944 human lung carcinoma cell line and Calu-6 human lung carcinoma cell line
    Species: Human
    Verified Customer | Posted 11/14/2017
    Human SSBP1 Antibody AF6588

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