Human/Mouse/Rat GAPDH Antibody

R&D Systems | Catalog # 2275-PC-100

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Canine, Xenograft

Applications

Validated:

Western Blot, Simple Western, Immunoprecipitation

Cited:

Western Blot, Western Blot Control

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
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Product Specifications

Immunogen

Synthetic peptide corresponding to a portion of the human GAPDH/G3PDH sequence

Specificity

Detects human, mouse, and rat GAPDH/G3PDH in Western blots.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat GAPDH Antibody

Detection of Human, Mouse, and Rat GAPDH/G3PDH antibody by Western Blot.

Detection of Human, Mouse, and Rat GAPDH/G3PDH by Western Blot.

Western blot shows lysates of U2OS human osteosarcoma cell line, A20 mouse B cell lymphoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat GAPDH Polyclonal Antibody (Catalog # 2275-PC) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for GAPDH at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human GAPDH by Simple WesternTM.

Simple Western shows lysates of Exosome Standards (LNCaP) (NBP3-11687), Exosome Standards (HT‑29) (NBP3-11685) and Jurkat human acute T cell leukemia cell line, loaded at 0.5 mg/ml. A specific band was detected for GAPDH at approximately 41 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Detection of Human, Mouse and Rat GAPDH by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, C2C12 mouse myoblast cell line, and C6 rat glioma cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH at approximately 43 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Mouse Human/Mouse/Rat GAPDH Antibody by Western Blot

Detection of Mouse Human/Mouse/Rat GAPDH Antibody by Western Blot

Higher levels of CD4 are found in macrophages expressing LCK.A) Schematic diagram of self-inactivating lentiviral construct used to express wild-type Lck and puroR (LCKWT P). A similar construct was used to express a kinase inactive form of Lck and puroR (LCKINACTIVE P) or to express puroR only (‘control P’). B) Detection of protein expression of LCK and CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-LCK and anti-CD4 antibodies. The loading control GAPDH was detected using anti-GAPDH antibody. C) Protein levels were measured with Odyssey software (Li-COR) and CD4 expression was normalised to GAPDH expression. Symbols represent normalised CD4 expression, relative to the PSC-macrophages control group, of two independent experiments. D) Detection of surface CD4 and total LCK expression. Representative two-colour immunofluorescence (dot plot) analysis is shown. Gates were determined by using the two relevant isotype control antibodies. Quantification of total LCK expression (E) and surface CD4 expression (F), expressed as the ratio of the geometric mean fluorescence intensity (MFI) over the isotype control ±SEM of independent experiments (n = 7). G) Detergent resistance of CD4, expressed as mean Flow Cytometric Detergent Resistance (FCDR) index of CD4 (n = 4) in PSC-macrophages, calculated as described in materials and methods. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Human/Mouse/Rat GAPDH Antibody by Western Blot

Detection of Mouse Human/Mouse/Rat GAPDH Antibody by Western Blot

CD4 knock-down in genetically modified stem cell-derived macrophages. C) Detection of protein expression of CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-CD4 antibodies. GAPDH, a loading control, was detected using anti-GAPDH antibody. Representative blot is shown. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GAPDH by Western Blot

Detection of Mouse GAPDH by Western Blot

Higher levels of CD4 are found in macrophages expressing LCK.A) Schematic diagram of self-inactivating lentiviral construct used to express wild-type Lck and puroR (LCKWT P). A similar construct was used to express a kinase inactive form of Lck and puroR (LCKINACTIVE P) or to express puroR only (‘control P’). B) Detection of protein expression of LCK and CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-LCK and anti-CD4 antibodies. The loading control GAPDH was detected using anti-GAPDH antibody. C) Protein levels were measured with Odyssey software (Li-COR) and CD4 expression was normalised to GAPDH expression. Symbols represent normalised CD4 expression, relative to the PSC-macrophages control group, of two independent experiments. D) Detection of surface CD4 and total LCK expression. Representative two-colour immunofluorescence (dot plot) analysis is shown. Gates were determined by using the two relevant isotype control antibodies. Quantification of total LCK expression (E) and surface CD4 expression (F), expressed as the ratio of the geometric mean fluorescence intensity (MFI) over the isotype control ±SEM of independent experiments (n = 7). G) Detergent resistance of CD4, expressed as mean Flow Cytometric Detergent Resistance (FCDR) index of CD4 (n = 4) in PSC-macrophages, calculated as described in materials and methods. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GAPDH by Western Blot

Detection of Mouse GAPDH by Western Blot

CD4 knock-down in genetically modified stem cell-derived macrophages.C) Detection of protein expression of CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-CD4 antibodies. GAPDH, a loading control, was detected using anti-GAPDH antibody. Representative blot is shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GAPDH by Western Blot

Detection of Mouse GAPDH by Western Blot

Higher levels of CD4 are found in macrophages expressing LCK.A) Schematic diagram of self-inactivating lentiviral construct used to express wild-type Lck and puroR (LCKWT P). A similar construct was used to express a kinase inactive form of Lck and puroR (LCKINACTIVE P) or to express puroR only (‘control P’). B) Detection of protein expression of LCK and CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-LCK and anti-CD4 antibodies. The loading control GAPDH was detected using anti-GAPDH antibody. C) Protein levels were measured with Odyssey software (Li-COR) and CD4 expression was normalised to GAPDH expression. Symbols represent normalised CD4 expression, relative to the PSC-macrophages control group, of two independent experiments. D) Detection of surface CD4 and total LCK expression. Representative two-colour immunofluorescence (dot plot) analysis is shown. Gates were determined by using the two relevant isotype control antibodies. Quantification of total LCK expression (E) and surface CD4 expression (F), expressed as the ratio of the geometric mean fluorescence intensity (MFI) over the isotype control ±SEM of independent experiments (n = 7). G) Detergent resistance of CD4, expressed as mean Flow Cytometric Detergent Resistance (FCDR) index of CD4 (n = 4) in PSC-macrophages, calculated as described in materials and methods. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat GAPDH Antibody

Application
Recommended Usage

Immunoprecipitation

1:1000-1:5000 dilution
Sample: L292 human fibrosarcoma cell line, WEHI 7.1 mouse T cell lymphoma cell line, and INT 407 human intestinal epithelial cell line

Simple Western

1-5 µg/mL
Sample: Exosome Standards (LNCap) (Catalog # NBP3-11687), Exosome Standards (HT-29) (Catalog # NBP3-11685), Jurkat human acute T cell leukemia cell line, C2C12 mouse myoblast cell line, and C6 rat glioma cell line

Western Blot

0.1 µg/mL
Sample: U2OS human osteosarcoma cell line, A20 mouse B cell lymphoma cell line, and L6 rat myoblast cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

Supplied as a solution containg PBS.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it 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 opening.
  • 6 months, -20 to -70 °C under sterile conditions after opening.

 

Background: GAPDH

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a 36‑40 kDa member of the GAPDH family of enzymes. It is a widely expressed heterotetramer that is found in both the nucleus and cytoplasm. Although GAPDH was initially identified as a glycolytic enzyme that converted G3P into 1,3 diphosphoglycerate, it is now recognized to participate in no less than endocytosis, membrane fusion, vesicular secretory transport, DNA replication and repair, and apoptosis. Human GAPDH is 335 amino acids (aa) in length and contains two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151‑155. GAPDH contains more than 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over amino acid 1‑150, human GAPDH shares 92% aa identity with mouse GAPDH.

Long Name

Glyceraldehyde-3-phosphate Dehydrogenase

Alternate Names

G3PDH

Entrez Gene IDs

2597 (Human); 14433 (Mouse); 24383 (Rat)

Gene Symbol

GAPDH

Additional GAPDH Products

Product Documents for Human/Mouse/Rat GAPDH Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat GAPDH Antibody

For research use only

Citations for Human/Mouse/Rat GAPDH Antibody

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